US20110119791A1

US20110119791A1 - Isolated polypeptides, polynucleotides useful for modifying water user efficiency, fertilizer use efficiency, biotic/abiotic stress tolerance, yield and biomass in plants

Info

Publication number: US20110119791A1
Application number: US12/810,855
Authority: US
Inventors: Gil Ronen; Basia J. Vinocur; Alex Diber; Sharon Ayal; Hagai Karchi; Yoav Herschkovitz
Original assignee: Evogene Ltd
Current assignee: Evogene Ltd
Priority date: 2007-12-27
Filing date: 2008-12-23
Publication date: 2011-05-19
Also published as: CA2709517A1; US20120222169A1; US9670501B2; AU2008344935C1; MX340504B; CA2709517C; WO2009083958A2; US8426682B2; CN101977928B; CN101977928A; WO2009083958A3; US20130219562A1; MX357387B; EP2240510B1; AU2008344935A1; WO2009083958A8; US10407690B2; EP2240510A2; ZA201004460B; AU2008344935B2

Abstract

Polynucleotides, polypeptides, plant cells expressing same and methods of using same for increasing abiotic stress tolerance water use efficiency (WUE), fertilizer use efficiency (FUE), biomass, vigor and/or yield of a plant. The method is effected by expressing within the plant an exogenous polynucleotide encoding a polypeptide comprising an amino acid sequence at least 80% homologous to the amino acid sequence selected from the group consisting of SEQ ID NOs:33, 34, 30, 27-29, 31, 32, 35-52, 1401-1403, 1405-1435, 1437-1494, 1496-1542, 1544-1553, 1555-1559, 1561-1827, 1829-1866, 1868-2450, 2453-2458, 2460-2463, 2465-2481, 2483, 2485-2746, 2765-2769, 3052-3065 and 3067-3259, thereby increasing the water use efficiency (WUE), the fertilizer use efficiency (FUE), the biomass, the vigor and/or the yield of the plant.

Description

FIELD AND BACKGROUND OF THE INVENTION

The present invention, in some embodiments thereof, relates to novel aquaporin polynucleotides and polypeptides, and more particularly, but not exclusively, to methods of using same for increasing abiotic stress tolerance, water use efficiency (WUE), fertilizer use efficiency (FUE), biomass, vigor and/or yield of a plant.
Abiotic stress conditions such as salinity, drought, flood, suboptimal temperature and toxic chemical pollution, cause substantial damage to agricultural plants. Most plants have evolved strategies to protect themselves against these conditions. However, if the severity and duration of the stress conditions are too great, the effects on plant development, growth and yield of most crop plants are profound. Furthermore, most of the crop plants are highly susceptible to abiotic stress (ABS) and thus necessitate optimal growth conditions for commercial crop yields. Continuous exposure to stress causes major alterations in the plant metabolism which ultimately leads to cell death and consequently yield losses.
The global shortage of water supply is one of the most severe agricultural problems affecting plant growth and crop yield and efforts are made to mitigate the harmful effects of desertification and salinization of the world's arable land. Thus, Agbiotech companies attempt to create new crop varieties which are tolerant to different abiotic stresses focusing mainly in developing new varieties that can tolerate water shortage for longer periods.
Studies have shown that plant adaptations to adverse environmental conditions are complex genetic traits with polygenic nature. When water supply is limited, the plant WUE is critical for the survival and yield of crop. Since water scarcity is increasing and water quality is reducing worldwide it is important to increase water productivity and plant WUE. Many of the environmental abiotic stresses, such as drought, low temperature or high salt, decrease root hydraulic conductance, affect plant growth and decrease crop productivity.
Genetic improvement of FUE in plants can be generated either via traditional breeding or via genetic engineering. Attempts to improve FUE in transgenic plants are described in U.S. Patent Applications 20020046419 to Choo, et al.; U.S. Pat. Appl. 20030233670 to Edgerton et al.; U.S. Pat. Appl. 20060179511 to Chomet et al.; Yanagisawa et al. [Proc. Natl. Acad. Sci. U.S.A. 2004, 101(20):7833-8]; Good A G et al. [Trends Plant Sci. 2004, 9(12):597-605]; and U.S. Pat. No. 6,084,153 to Good et al.
Aquaporins (AQPs), the water channel proteins, are involved in transport of water through the membranes, maintenance of cell water balance and homeostasis under changing environmental and developmental conditions [Maurel C. Plant aquaporins: Novel functions and regulation properties. FEBS Lett. 2007, 581(12):2227-36]. These proteins are considered to be the main passage enabling transport of water and small neutral solutes such as urea and CO₂through the membrane [Maurel C. Plant aquaporins: Novel functions and regulation properties. FEBS Lett. 2007 Jun. 12; 581(12):2227-36]. In plants, AQPs are present as four subfamilies of intrinsic proteins: plasma membrane (PIP), tonoplast (TIP), small and basic (SIP) and NOD26-like (NIP). The total number of AQP members in plants, as compared to animals, appears to be surprisingly high [Maurel C., 2007 (Supra)]. For instance, 35 AQP genes have been identified in the Arabidopsis genome [Quigley F, et al., “From genome to function: the Arabidopsis aquaporins”. Genome Biol. 2002, 3(1):RESEARCH0001.1-1.17], 36 in maize [Chaumont F, et al., 2001, “Aquaporins constitute a large and highly divergent protein family in maize. Plant Physiol”, 125(3):1206-15], and 33 in rice [Sakurai, J., et a., 2005, Identification of 33 rice aquaporin genes and analysis of their expression and function. Plant Cell Physiol. 46, 1568-1577]. The high number of AQPs in plants suggests a diverse role and differential regulation under variable environmental conditions [Maurel C., 2007 (Supra)].
WO2004/104162 to the present inventors teaches polynucleotide sequences and methods of utilizing same for increasing the tolerance of a plant to abiotic stresses and/or increasing the biomass of a plant.
WO2007/020638 to the present inventors teaches polynucleotide sequences and methods of utilizing same for increasing the tolerance of a plant to abiotic stresses and/or increasing the biomass, vigor and/or yield of a plant.
Lian H L, et al., 2006 (Cell Res. 16: 651-60) over-expressed members of the PIP1 subgroup of AQPs in rice. Aharon R., et al. 2003 (Plant Cell, 15: 439-47) over-expressed the Arabidopsis plasma membrane aquaporin, PIP1b, in transgenic tobacco plants.

SUMMARY OF THE INVENTION

According to an aspect of some embodiments of the present invention there is provided a method of increasing abiotic stress tolerance of a plant, comprising expressing within the plant an exogenous polynucleotide encoding a polypeptide comprising an amino acid sequence at least 80% homologous to the amino acid sequence selected from the group consisting of SEQ ID NOs: 33, 34, 30, 27-29, 31, 32, 35-52, 1401-1403, 1405-1435, 1437-1494, 1496-1542, 1544-1553, 1555-1559, 1561-1827, 1829-1866, 1868-2450, 2453-2458, 2460-2463, 2465-2481, 2483, 2485-2746, 2765-2769, 3052-3065 and 3067-3259, thereby increasing the abiotic stress tolerance of the plant.
According to an aspect of some embodiments of the present invention there is provided a method of increasing water use efficiency (WUE), fertilizer use efficiency (FUE), biomass, vigor and/or yield of a plant, comprising expressing within the plant an exogenous polynucleotide encoding a polypeptide comprising an amino acid sequence at least 80% homologous to the amino acid sequence selected from the group consisting of SEQ ID NOs:33, 34, 30, 27-29, 31, 32, 35-52, 1401-1403, 1405-1435, 1437-1494, 1496-1542, 1544-1553, 1555-1559, 1561-1827, 1829-1866, 1868-2450, 2453-2458, 2460-2463, 2465-2481, 2483, 2485-2746, 2765-2769, 3052-3065 and 3067-3259, thereby increasing the water use efficiency (WUE), the fertilizer use efficiency (FUE), the biomass, the vigor and/or the yield of the plant.
According to an aspect of some embodiments of the present invention there is provided an isolated polynucleotide comprising a nucleic acid sequence at least 80% identical to the nucleic acid sequence selected from the group consisting of SEQ ID NOs:7, 8, 4, 1-3, 5, 6, 9-26, 53-55, 57-87, 89-147, 149-195, 197-206, 208-212, 214-480, 482-519, 521-1103, 1106-1111, 1113-1116, 1118-1134, 1136, 1138-1400, 2748-2764, 2843-2857 and 2859-3051.
According to an aspect of some embodiments of the present invention there is provided a nucleic acid construct, comprising the isolated polynucleotide of the invention and a promoter for directing transcription of the nucleic acid sequence.
According to an aspect of some embodiments of the present invention there is provided an isolated polypeptide, comprising an amino acid sequence at least 80% homologous to the amino acid sequence selected from the group consisting of SEQ ID NOs:33, 34, 30, 27-29, 31, 32, 35-52, 1401-1403, 1405-1435, 1437-1494, 1496-1542, 1544-1553, 1555-1559, 1561-1827, 1829-1866, 1868-2450, 2453-2458, 2460-2463, 2465-2481, 2483, 2485-2746, 2765-2769, 3052-3065 and 3067-3259.
According to an aspect of some embodiments of the present invention there is provided a plant cell comprising an exogenous polypeptide having an amino acid sequence at least 80% homologous to the amino acid sequence selected from the group consisting of SEQ ID NOs:33, 34, 30, 27-29, 31, 32, 35-52, 1401-1403, 1405-1435, 1437-1494, 1496-1542, 1544-1553, 1555-1559, 1561-1827, 1829-1866, 1868-2450, 2453-2458, 2460-2463, 2465-2481, 2483, 2485-2746, 2765-2769, 3052-3065 and 3067-3259.
According to an aspect of some embodiments of the present invention there is provided a plant cell comprising an exogenous polynucleotide comprising a nucleic acid sequence at least 80% homologous to the nucleic acid sequence selected from the group consisting of SEQ ID NOs:7, 8, 4, 1-3, 5, 6, 9-26, 53-55, 57-87, 89-147, 149-195, 197-206, 208-212, 214-480, 482-519, 521-1103, 1106-1111, 1113-1116, 1118-1134, 1136, 1138-1400, 2748-2764, 2843-2857 and 2859-3051.
According to an aspect of some embodiments of the present invention there is provided a method of increasing abiotic stress tolerance of a plant, comprising expressing within the plant an exogenous polynucleotide encoding a polypeptide comprising the amino acid sequence set forth by SEQ ID NO:33, 34, 30, 27-29, 31, 32, 35-52, 1401-1403, 1405-1435, 1437-1494, 1496-1542, 1544-1553, 1555-1559, 1561-1827, 1829-1866, 1868-2450, 2453-2458, 2460-2463, 2465-2481, 2483, 2485-2746, 2765-2769, 3052-3065, 3067-3258 or 3259, thereby increasing the abiotic stress tolerance of the plant.
According to an aspect of some embodiments of the present invention there is provided a method of increasing water use efficiency (WUE), fertilizer use efficiency (FUE), biomass, vigor and/or yield of a plant, comprising expressing within the plant an exogenous polynucleotide encoding a polypeptide comprising the amino acid sequence set forth by SEQ ID NO:33, 34, 30, 27-29, 31, 32, 35-52, 1401-1403, 1405-1435, 1437-1494, 1496-1542, 1544-1553, 1555-1559, 1561-1827, 1829-1866, 1868-2450, 2453-2458, 2460-2463, 2465-2481, 2483, 2485-2746, 2765-2769, 3052-3065, 3067-3258 or 3259, thereby increasing the water use efficiency (WUE), the fertilizer use efficiency (FUE), the biomass, the vigor and/or the yield of the plant.
According to an aspect of some embodiments of the present invention there is provided an isolated polynucleotide comprising the nucleic acid sequence set forth by SEQ ID NO:7, 8, 4, 1-3, 5, 6, 9-26, 53-55, 57-87, 89-147, 149-195, 197-206, 208-212, 214-480, 482-519, 521-1103, 1106-1111, 1113-1116, 1118-1134, 1136, 1138-1400, 2748-2764, 2843-2857, 2859-3050 or 3051.
According to an aspect of some embodiments of the present invention there is provided a nucleic acid construct, comprising the isolated polynucleotide of the invention and a promoter for directing transcription of the nucleic acid sequence.
According to an aspect of some embodiments of the present invention there is provided an isolated polypeptide, comprising the amino acid sequence set forth by SEQ ID NO:33, 34, 30, 27-29, 31, 32, 35-52, 1401-1403, 1405-1435, 1437-1494, 1496-1542, 1544-1553, 1555-1559, 1561-1827, 1829-1866, 1868-2450, 2453-2458, 2460-2463, 2465-2481, 2483, 2485-2746, 2765-2769, 3052-3065, 3067-3258 or 3259.
According to an aspect of some embodiments of the present invention there is provided a plant cell comprising an exogenous polypeptide having the amino acid sequence set forth by SEQ ID NO:33, 34, 30, 27-29, 31, 32, 35-52, 1401-1403, 1405-1435, 1437-1494, 1496-1542, 1544-1553, 1555-1559, 1561-1827, 1829-1866, 1868-2450, 2453-2458, 2460-2463, 2465-2481, 2483, 2485-2746, 2765-2769, 3052-3065, 3067-3258 or 3259.
According to an aspect of some embodiments of the present invention there is provided a plant cell comprising an exogenous polynucleotide comprising the nucleic acid sequence set forth by SEQ ID NO:7, 8, 4, 1-3, 5, 6, 9-26, 53-55, 57-87, 89-147, 149-195, 197-206, 208-212, 214-480, 482-519, 521-1103, 1106-1111, 1113-1116, 1118-1134, 1136, 1138-1400, 2748-2764, 2843-2857, 2859-3050 or 3051.
According to some embodiments of the invention, the polynucleotide is selected from the group consisting of SEQ ID NOs:7, 8, 4, 1-3, 5, 6, 9-26, 53-55, 57-87, 89-147, 149-195, 197-206, 208-212, 214-480, 482-519, 521-1103, 1106-1111, 1113-1116, 1118-1134, 1136, 1138-1400, 2748-2764, 2843-2857 and 2859-3051.
According to some embodiments of the invention, the amino acid sequence is selected from the group consisting of SEQ ID NOs:33, 34, 30, 27-29, 31, 32, 35-52, 1401-1403, 1405-1435, 1437-1494, 1496-1542, 1544-1553, 1555-1559, 1561-1827, 1829-1866, 1868-2450, 2453-2458, 2460-2463, 2465-2481, 2483, 2485-2746, 2765-2769, 3052-3065 and 3067-3259.
According to some embodiments of the invention, the polypeptide is selected from the group consisting of SEQ ID NOs:33, 34, 30, 27-29, 31, 32, 35-52, 1401-1403, 1405-1435, 1437-1494, 1496-1542, 1544-1553, 1555-1559, 1561-1827, 1829-1866, 1868-2450, 2453-2458, 2460-2463, 2465-2481, 2483, 2485-2746, 2765-2769, 3052-3065 and 3067-3259.
According to some embodiments of the invention, the abiotic stress is selected from the group consisting of salinity, water deprivation, low temperature, high temperature, heavy metal toxicity, anaerobiosis, nutrient deficiency, nutrient excess, atmospheric pollution and UV irradiation.
According to some embodiments of the invention, the method further comprising growing the plant expressing the exogenous polynucleotide under the abiotic stress.
According to some embodiments of the invention, the promoter is a constitutive promoter.
According to some embodiments of the invention, the plant cell forms a part of a plant.
Unless otherwise defined, all technical and/or scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which the invention pertains. Although methods and materials similar or equivalent to those described herein can be used in the practice or testing of embodiments of the invention, exemplary methods and/or materials are described below. In case of conflict, the patent specification, including definitions, will control. In addition, the materials, methods, and examples are illustrative only and are not intended to be necessarily limiting.

BRIEF DESCRIPTION OF THE DRAWINGS

Some embodiments of the invention are herein described, by way of example only, with reference to the accompanying drawings. With specific reference now to the drawings in detail, it is stressed that the particulars shown are by way of example and for purposes of illustrative discussion of embodiments of the invention. In this regard, the description taken with the drawings makes apparent to those skilled in the art how embodiments of the invention may be practiced.

In the drawings:

FIG. 1 is a schematic illustration of the pGI binary plasmid used for expressing the isolated polynucleotide sequences of the invention. RB—T-DNA right border; LB—T-DNA left border; H—HindIII restriction enzyme; X—XbaI restriction enzyme; B—BamHI restriction enzyme; S—SalI restriction enzyme; Sm—SmaI restriction enzyme; R-I—EcoRI restriction enzyme; Sc—SacI/SstI/Ecl136II; (numbers)—Length in base-pairs; NOS pro=nopaline synthase promoter; NPT-II=neomycin phosphotransferase gene; NOS ter=nopaline synthase terminator; Poly-A signal (polyadenylation signal); GUSintron—the GUS reporter gene (coding sequence and intron). The isolated polynucleotide sequences of some embodiments of the invention were cloned into the vector while replacing the GUSintron reporter gene.

FIGS. 2A-B are images depicting root development of plants grown in transparent agar plates. The different transgenes were grown in transparent agar plates for 10-15 days and the plates were photographed every 2-5 days starting at day 1. FIG. 2A—An exemplary image of plants taken following 12 days on agar plates. FIG. 2B—An exemplary image of root analysis in which the length of the root measured is represented by a red arrow.

FIGS. 3A-F are histograms depicting the total economic fruit yield, plant biomass and harvest index for TOM-ABST36 (black bar) vs. control (white bar) plants growing in the commercial greenhouse under a 200 mM sodium chloride (NaCl) irrigation regime (FIG. 3A-C, respectively), or under two different water-stress regimes (WLI-1 and WLI-2; FIG. 3D-F, respectively). Yield performance was compared to plants growing under standard irrigation conditions (0 mM NaCl and WLI-0). Results are the average of the four independent events. *Significantly different at P≦0.05.

FIGS. 3G-J are photographs of transgenic tomato plants or control plants grown under various conditions. FIG. 3G—TOM-ABST36 plants growing under regular irrigation conditions; FIG. 3H—control plants growing under regular irrigation conditions; FIG. 3I—TOM-ABST36 plants after growing under a 200-mM NaCl-irrigation regime during the entire growing season; FIG. 3J—control plants after growing under a 200-mM NaCl-irrigation regime during the entire growing season.

DESCRIPTION OF SPECIFIC EMBODIMENTS OF THE INVENTION

The present invention, in some embodiments thereof, relates to novel aquaporin polynucleotides and polypeptides, and more particularly, but not exclusively, to methods of using same for increasing abiotic stress tolerance, water use efficiency, fertilizer use efficiency, biomass, vigor and/or yield of a plant.
Before explaining at least one embodiment of the invention in detail, it is to be understood that the invention is not necessarily limited in its application to the details set forth in the following description or exemplified by the Examples. The invention is capable of other embodiments or of being practiced or carried out in various ways.
While reducing the invention to practice, the present inventors have identified novel aquaporin (AQP) polynucleotides and polypeptides encoded thereby.
Thus, as shown in the Examples section which follows, the present inventors have employed a bioinformatics approach which combines digital expression analysis and cross-species comparative genomics and screened 7.2 million expressed sequence tags (ESTs) from 1,195 relevant EST's libraries of both monocot and dicot plant species. Using this approach 1,114 different AQP genes have been identified and were further classified to 11 subgroups (Table 1). Further analysis revealed that ESTs of the TIP2 subgroup are significantly over-represented in both plants' roots and in plants exposed to abiotic stress (ABS), and that polypeptides (e.g., SEQ ID NOs: 27-28, 45-48, Table 2) encoded by polynucleotides of the TIP2 subgroup (e.g., SEQ ID NOs:1, 2, 19-22, Table 2) share a common consensus sequence TLXFXFAGVGS (SEQ ID NO:2826). Based on over-representation in roots, ABS conditions and tissues with low water levels (such as seed and pollen) additional polynucleotides of the aquaporin gene family were identified (SEQ ID NOs: 3-18, 23-26, Table 2), as well as homologues or orthologues thereof (SEQ ID NOs:53-1400, 2844-3051 for polynucleotides and SEQ ID NOs:1401-2746, 3052-3259 for polypeptides; Table 3). Moreover, quantitative RT-PCR analysis demonstrated increased expression of representative AQP genes (e.g., SEQ ID NOs:5, 6 and 7) under salt stress, which was higher in plants exhibiting salt tolerance as compared to plants which are sensitive to salt stress (Table 5, Example 2 of the Examples section which follows). As is further described in Examples 3-4 of the Examples section which follows, representative AQP polynucleotides were cloned (Tables 7, 8 and 9) and transgenic plants over-expressing same were generated (Example 4). These plants were shown to exhibit increased tolerance to various abiotic stresses such as osmotic stress (Tables 10-14; Example 5) and salinity stress (Tables 30-45; Example 6), increased fertilizer use efficiency (under nitrogen limiting conditions, Tables 59-68, Example 7) and increased growth, biomass and yield under normal [Tables 15-29 (Example 5), 46-58 (Example 6)] or abiotic stress conditions conditions (Examples 5-8). Altogether, these results suggest the use of the AQP polynucleotides and polypeptides of the invention for increasing abiotic stress tolerance, water use efficiency, fertilizer use efficiency, biomass, vigor and/or yield of a plant.
It should be noted that polypeptides or polynucleotides which affect (e.g., increase) plant metabolism, growth, reproduction and/or viability under stress, can also affect the plant growth, biomass, yield and/or vigor under optimal conditions.
Thus, according to one aspect of the invention, there is provided a method of increasing abiotic stress tolerance, water use efficiency, fertilizer use efficiency, growth, biomass, yield and/or vigor of a plant. The method is effected by expressing within the plant an exogenous polynucleotide encoding a polypeptide comprising the amino acid consensus sequence TLXFXFAGVGS as set forth by SEQ ID NO:2826, wherein expression of the polypeptide promotes plants' biomass/vigor and/or yield under normal or stress conditions.
It is suggested that the polypeptide's activity is structurally associated with the integrity of the above consensus sequence (SEQ ID NO:2826). In some embodiments of this aspect of the present invention, the activity is a water channel activity which typically resides in the vacuaolar membrane (tonoplast) and/or the plasma membrane of the plant cell and enables the transport of water and/or small neutral solutes such as urea, nitrates and carbon dioxide (CO₂) through the membrane.
The phrase “abiotic stress” as used herein refers to any adverse effect on metabolism, growth, reproduction and/or viability of a plant. Accordingly, abiotic stress can be induced by suboptimal environmental growth conditions such as, for example, salinity, water deprivation, flooding, freezing, low or high temperature, heavy metal toxicity, anaerobiosis, nutrient deficiency, atmospheric pollution or UV irradiation. The implications of abiotic stress are discussed in the Background section.
The phrase “abiotic stress tolerance” as used herein refers to the ability of a plant to endure an abiotic stress without suffering a substantial alteration in metabolism, growth, productivity and/or viability.
As used herein the phrase “water use efficiency (WUE)” refers to the level of organic matter produced per unit of water consumed by the plant, i.e., the dry weight of a plant in relation to the plant's water use, e.g., the biomass produced per unit transpiration.
As used herein the phrase “fertilizer use efficiency” refers to the uptake, spread, absorbent, accumulation, relocation (within the plant) and use of one or more of the minerals and organic moieties absorbed from the soil, such as nitrogen, phosphates and/or potassium.
As used herein the phrase “plant biomass” refers to the amount (measured in grams of air-dry tissue) of a tissue produced from the plant in a growing season, which could also determine or affect the plant yield or the yield per growing area.
As used herein the phrase “plant yield” refers to the amount (as determined by weight/size) or quantity (numbers) of tissue produced per plant or per growing season. Hence increased yield could affect the economic benefit one can obtain from the plant in a certain growing area and/or growing time.
As used herein the phrase “plant vigor” refers to the amount (measured by weight) of tissue produced by the plant in a given time. Hence increase vigor could determine or affect the plant yield or the yield per growing time or growing area.
As used herein the term “increasing” refers to at least about 2%, at least about 3%, at least about 4%, at least about 5%, at least about 10%, at least about 15%, at least about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, increase in plant abiotic stress tolerance, water use efficiency, fertilizer use efficiency, growth, biomass, yield and/or vigor as compared to a native plant [i.e., a plant not modified with the biomolecules (polynucleotide or polypeptides) of the invention, e.g., a non-transformed plant of the same species which is grown under the same growth conditions).
As used herein, the phrase “exogenous polynucleotide” refers to a heterologous nucleic acid sequence which may not be naturally expressed within the plant or which overexpression in the plant is desired. The exogenous polynucleotide may be introduced into the plant in a stable or transient manner, so as to produce a ribonucleic acid (RNA) molecule and/or a polypeptide molecule. It should be noted that the exogenous polynucleotide may comprise a nucleic acid sequence which is identical or partially homologous to an endogenous nucleic acid sequence of the plant.
According to some embodiments of the invention, the exogenous polynucleotide of the invention encodes a polypeptide having an amino acid sequence at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 81%, at least about 82%, at least about 83%, at least about 84%, at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or more say 100% homologous to the amino acid sequence selected from the group consisting of SEQ ID NOs: 27-28, 45-48, 1401-1403, 1405-1435, 1437-1494, 1496-1542, 1544-1553, 1555-1559, 1561, 2449-2450, 2453-2458, 2460-2463, 2465-2481, 2483, 2484 and 2765.
Homology (e.g., percent homology) can be determined using any homology comparison software, including for example, the BlastP or TBLASTN software of the National Center of Biotechnology Information (NCBI) such as by using default parameters, when starting from a polypeptide sequence; or the tBLASTX algorithm (available via the NCBI) such as by using default parameters, which compares the six-frame conceptual translation products of a nucleotide query sequence (both strands) against a protein sequence database.
Homologous sequences include both orthologous and paralogous sequences. The term “paralogous” relates to gene-duplications within the genome of a species leading to paralogous genes. The term “orthologous” relates to homologous genes in different organisms due to ancestral relationship.
One option to identify orthologues in monocot plant species is by performing a reciprocal blast search. This may be done by a first blast involving blasting the sequence-of-interest against any sequence database, such as the publicly available NCBI database which may be found at: Hypertext Transfer Protocol://World Wide Web (dot) ncbi (dot) nlm (dot) nih (dot) gov. If orthologues in rice were sought, the sequence-of-interest would be blasted against, for example, the 28,469 full-length cDNA clones from Oryza sativa Nipponbare available at NCBI. The blast results may be filtered. The full-length sequences of either the filtered results or the non-filtered results are then blasted back (second blast) against the sequences of the organism from which the sequence-of-interest is derived. The results of the first and second blasts are then compared. An orthologue is identified when the sequence resulting in the highest score (best hit) in the first blast identifies in the second blast the query sequence (the original sequence-of-interest) as the best hit. Using the same rational a paralogue (homolog to a gene in the same organism) is found. In case of large sequence families, the ClustalW program may be used [Hypertext Transfer Protocol://World Wide Web (dot) ebi (dot) ac (dot) uk/Tools/clustalw2/index (dot) html], followed by a neighbor-joining tree (Hypertext Transfer Protocol://en (dot) wikipedia (dot) org/wiki/Neighbor-joining) which helps visualizing the clustering.
According to some embodiments of the invention, the exogenous polynucleotide encodes a polypeptide consisting of the amino acid sequence set forth by SEQ ID NO:27-28, 45-48, 1401-1403, 1405-1435, 1437-1494, 1496-1542, 1544-1553, 1555-1559, 1561, 2449-2450, 2453-2458, 2460-2463, 2465-2481, 2483, 2484 or 2765.
According to some embodiments of the invention the exogenous polynucleotide comprises a nucleic acid sequence which is at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 81%, at least about 82%, at least about 83%, at least about 84%, at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, e.g., 100% identical to the nucleic acid sequence selected from the group consisting of SEQ ID NOs:1, 2, 19, 20-22, 53-55, 57-87, 89-141, 143-147, 149-195, 197-206, 208-212, 214, 1102-1103, 1106-1111, 1113-1116, 1118-1134, 1136, 2751-2752 and 2748-2750.
Identity (e.g., percent homology) can be determined using any homology comparison software, including for example, the BlastN software of the National Center of Biotechnology Information (NCBI) such as by using default parameters.
According to some embodiments of the invention the exogenous polynucleotide is set forth by SEQ ID NO:1, 2, 19, 20-22, 53-55, 57-87, 89-141, 143-147, 149-195, 197-206, 208-212, 214, 1102-1103, 1106-1111, 1113-1116, 1118-1134, 1136, 2751-2752, 2748-2749, or 2750.
Notwithstanding the above, additional AQP polynucleotides and polypeptides encoded thereby are contemplated by the present teachings.
According to some embodiments of the invention, the exogenous polynucleotide encodes a polypeptide having an amino acid sequence at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, e.g., 100% homologous to SEQ ID NO:33, 34, 30, 27-29, 31, 32, 35-52, 1401-1403, 1405-1435, 1437-1494, 1496-1542, 1544-1553, 1555-1559, 1561-1827, 1829-1866, 1868-2450, 2453-2458, 2460-2463, 2465-2481, 2483, 2485-2746, 2765-2769, 3052-3065, 3067-3258 or 3259.
According to some embodiments of the invention, the exogenous polynucleotide encodes a polypeptide consisting of the amino acid sequence set forth by SEQ ID NO:33, 34, 30, 27-29, 31, 32, 35-52, 1401-1403, 1405-1435, 1437-1494, 1496-1542, 1544-1553, 1555-1559, 1561-1827, 1829-1866, 1868-2450, 2453-2458, 2460-2463, 2465-2481, 2483, 2485-2746, 2765-2769, 3052-3065, 3067-3258 or 3259.
In an exemplary embodiment the exogenous polynucleotide does not encode a polypeptide having the amino acid sequence selected from the group consisting of SEQ ID NOs: 1828, 1867, 1404, 1436, 1495, 1543, 1554, 1560, 2451, 2452, 2459, 2464, 2482, 2484 and 3066.
According to some embodiments of the invention, the exogenous polynucleotide is at least at least about 60%, least at least about 65%, least at least about 70%, least at least about 75%least at least about 80%, at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, e.g., 100% identical to SEQ ID NO:7, 8, 4, 1-3, 5, 6, 9-26, 53-55, 57-87, 89-147, 149-195, 197-206, 208-212, 214-480, 482-519, 521-1103, 1106-1111, 1113-1116, 1118-1134, 1136, 1138-1400, 2748-2764, 2843-2857, 2859-3050 or 3051.
According to some embodiments of the invention, the polynucleotide is set forth by SEQ ID NO:7, 8, 4, 1-3, 5, 6, 9-26, 53-55, 57-87, 89-147, 149-195, 197-206, 208-212, 214-480, 482-519, 521-1103, 1106-1111, 1113-1116, 1118-1134, 1136, 1138-1400, 2748-2764, 2843-2857, 2859-3050 or 3051.
In an exemplary embodiments the exogenous polynucleotide is not the polynucleotide set forth by SEQ ID NO: 481, 520, 56, 88, 148, 196, 207, 213, 1104, 1105, 1112, 1117, 1135, 1137 or 2858.
As used herein the term “polynucleotide” refers to a single or double stranded nucleic acid sequence which is isolated and provided in the form of an RNA sequence, a complementary polynucleotide sequence (cDNA), a genomic polynucleotide sequence and/or a composite polynucleotide sequences (e.g., a combination of the above).
As used herein the phrase “complementary polynucleotide sequence” refers to a sequence, which results from reverse transcription of messenger RNA using a reverse transcriptase or any other RNA dependent DNA polymerase. Such a sequence can be subsequently amplified in vivo or in vitro using a DNA dependent DNA polymerase.
As used herein the phrase “genomic polynucleotide sequence” refers to a sequence derived (isolated) from a chromosome and thus it represents a contiguous portion of a chromosome.
As used herein the phrase “composite polynucleotide sequence” refers to a sequence, which is at least partially complementary and at least partially genomic. A composite sequence can include some exonal sequences required to encode the polypeptide of the present invention, as well as some intronic sequences interposing therebetween. The intronic sequences can be of any source, including of other genes, and typically will include conserved splicing signal sequences. Such intronic sequences may further include cis acting expression regulatory elements.
According to some embodiments of the invention, the polynucleotide of the invention comprises no more than 5000 nucleic acids in length. According to some embodiments of the invention, the polynucleotide of the invention comprises no more than 4000 nucleic acids in length, e.g., no more than 3000 nucleic acids, e.g., no more than 2500 nucleic acids.
Nucleic acid sequences encoding the polypeptides of the present invention may be optimized for expression. A non-limiting example of an optimized nucleic acid sequence is provided in SEQ ID NO:2751, which encodes an optimized polypeptide comprising the amino acid sequence set forth by SEQ ID NO:27. Examples of such sequence modifications include, but are not limited to, an altered G/C content to more closely approach that typically found in the plant species of interest, and the removal of codons atypically found in the plant species commonly referred to as codon optimization.
The phrase “codon optimization” refers to the selection of appropriate DNA nucleotides for use within a structural gene or fragment thereof that approaches codon usage within the plant of interest. Therefore, an optimized gene or nucleic acid sequence refers to a gene in which the nucleotide sequence of a native or naturally occurring gene has been modified in order to utilize statistically-preferred or statistically-favored codons within the plant. The nucleotide sequence typically is examined at the DNA level and the coding region optimized for expression in the plant species determined using any suitable procedure, for example as described in Sardana et al. (1996, Plant Cell Reports 15:677-681). In this method, the standard deviation of codon usage, a measure of codon usage bias, may be calculated by first finding the squared proportional deviation of usage of each codon of the native gene relative to that of highly expressed plant genes, followed by a calculation of the average squared deviation. The formula used is: 1 SDCU=n=1N[(Xn−Yn)/Yn]2/N, where Xn refers to the frequency of usage of codon n in highly expressed plant genes, where Yn to the frequency of usage of codon n in the gene of interest and N refers to the total number of codons in the gene of interest. A Table of codon usage from highly expressed genes of dicotyledonous plants is compiled using the data of Murray et al. (1989, Nuc Acids Res. 17:477-498).
One method of optimizing the nucleic acid sequence in accordance with the preferred codon usage for a particular plant cell type is based on the direct use, without performing any extra statistical calculations, of codon optimization Tables such as those provided on-line at the Codon Usage Database through the NIAS (National Institute of Agrobiological Sciences) DNA bank in Japan (Hypertext Transfer Protocol://World Wide Web (dot) kazusa (dot) or (dot) jp/codon/). The Codon Usage Database contains codon usage tables for a number of different species, with each codon usage Table having been statistically determined based on the data present in Genbank.
By using the above Tables to determine the most preferred or most favored codons for each amino acid in a particular species (for example, rice), a naturally-occurring nucleotide sequence encoding a protein of interest can be codon optimized for that particular plant species. This is effected by replacing codons that may have a low statistical incidence in the particular species genome with corresponding codons, in regard to an amino acid, that are statistically more favored. However, one or more less-favored codons may be selected to delete existing restriction sites, to create new ones at potentially useful junctions (5′ and 3′ ends to add signal peptide or termination cassettes, internal sites that might be used to cut and splice segments together to produce a correct full-length sequence), or to eliminate nucleotide sequences that may negatively effect mRNA stability or expression.
The naturally-occurring encoding nucleotide sequence may already, in advance of any modification, contain a number of codons that correspond to a statistically-favored codon in a particular plant species. Therefore, codon optimization of the native nucleotide sequence may comprise determining which codons, within the native nucleotide sequence, are not statistically-favored with regards to a particular plant, and modifying these codons in accordance with a codon usage table of the particular plant to produce a codon optimized derivative. A modified nucleotide sequence may be fully or partially optimized for plant codon usage provided that the protein encoded by the modified nucleotide sequence is produced at a level higher than the protein encoded by the corresponding naturally occurring or native gene. Construction of synthetic genes by altering the codon usage is described in for example PCT Patent Application 93/07278.
Thus, the invention encompasses nucleic acid sequences described hereinabove; fragments thereof, sequences hybridizable therewith, sequences homologous thereto, sequences encoding similar polypeptides with different codon usage, altered sequences characterized by mutations, such as deletion, insertion or substitution of one or more nucleotides, either naturally occurring or man induced, either randomly or in a targeted fashion.
As mentioned, the present inventors have uncovered previously uncharacterized polypeptides which share the amino acid consensus sequence set forth by SEQ ID NO:2826.
Thus, the invention provides an isolated polypeptide having an amino acid sequence at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 81%, at least about 82%, at least about 83%, at least about 84%, at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or more say 100% homologous to an amino acid sequence selected from the group consisting of SEQ ID NO: 27-28, 45-48, 1401-1403, 1405-1435, 1437-1494, 1496-1542, 1544-1553, 1555-1559, 1561, 2449-2450, 2453-2458, 2460-2463, 2465-2481, 2483, 2484 and 2765.
According to some embodiments of the invention, the invention provides an isolated polypeptide having an amino acid sequence at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 81%, at least about 82%, at least about 83%, at least about 84%, at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or more say 100% homologous to an amino acid sequence selected from the group consisting of SEQ ID NOs:33, 34, 30, 27-29, 31, 32, 35-52, 1401-1403, 1405-1435, 1437-1494, 1496-1542, 1544-1553, 1555-1559, 1561-1827, 1829-1866, 1868-2450, 2453-2458, 2460-2463, 2465-2481, 2483, 2485-2746, 2765-2769, 3052-3065 and 3067-3259.
According to some embodiments of the invention, the polypeptide is set forth by SEQ ID NO: 33, 34, 30, 27-29, 31, 32, 35-52, 1401-1403, 1405-1435, 1437-1494, 1496-1542, 1544-1553, 1555-1559, 1561-1827, 1829-1866, 1868-2450, 2453-2458, 2460-2463, 2465-2481, 2483, 2485-2746, 2765-2769, 3052-3065, 3067-3258 or 3259.
In an exemplary embodiment the polypeptide is not the polypeptide set forth by SEQ ID NO: 1828, 1867, 1404, 1436, 1495, 1543, 1554, 1560, 2451, 2452, 2459, 2464, 2482, 2484 or 3066.
The invention also encompasses fragments of the above described polypeptides and polypeptides having mutations, such as deletions, insertions or substitutions of one or more amino acids, either naturally occurring or man induced, either randomly or in a targeted fashion.
The term “plant” as used herein encompasses whole plants, ancestors and progeny of the plants and plant parts, including seeds, shoots, stems, roots (including tubers), and plant cells, tissues and organs. The plant may be in any form including suspension cultures, embryos, meristematic regions, callus tissue, leaves, gametophytes, sporophytes, pollen, and microspores. Plants that are particularly useful in the methods of the invention include all plants which belong to the superfamily Viridiplantae, in particular monocotyledonous and dicotyledonous plants including a fodder or forage legume, ornamental plant, food crop, tree, or shrub selected from the list comprising Acacia spp., Acer spp., Actinidia spp., Aesculus spp., Agathis australis, Albizia amara, Alsophila tricolor, Andropogon spp., Arachis spp, Areca catechu, Astelia fragrans, Astragalus cicer, Baikiaea plurijuga, Betula spp., Brassica spp., Bruguiera gymnorrhiza, Burkea africana, Butea frondosa, Cadaba farinosa, Calliandra spp, Camellia sinensis, Canna indica, Capsicum spp., Cassia spp., Centroema pubescens, Chacoomeles spp., Cinnamomum cassia, Coffea arabica, Colophospermum mopane, Coronillia varia, Cotoneaster serotina, Crataegus spp., Cucumis spp., Cupressus spp., Cyathea dealbata, Cydonia oblonga, Cryptomeria japonica, Cymbopogon spp., Cynthea dealbata, Cydonia oblonga, Dalbergia monetaria, Davallia divaricata, Desmodium spp., Dicksonia squarosa, Dibeteropogon amplectens, Dioclea spp, Dolichos spp., Dorycnium rectum, Echinochloa pyramidalis, Ehraffia spp., Eleusine coracana, Eragrestis spp., Erythrina spp., Eucalypfus spp., Euclea schimperi, Eulalia vi/losa, Pagopyrum spp., Feijoa sellowlana, Fragaria spp., Flemingia spp, Freycinetia banksli, Geranium thunbergii, GinAgo biloba, Glycine javanica, Gliricidia spp, Gossypium hirsutum, Grevillea spp., Guibourtia coleosperma, Hedysarum spp., Hemaffhia altissima, Heteropogon contoffus, Hordeum vulgare, Hyparrhenia rufa, Hypericum erectum, Hypeffhelia dissolute, Indigo incamata, Iris spp., Leptarrhena pyrolifolia, Lespediza spp., Lettuca spp., Leucaena leucocephala, Loudetia simplex, Lotonus bainesli, Lotus spp., Macrotyloma axillare, Malus spp., Manihot esculenta, Medicago saliva, Metasequoia glyptostroboides, Musa sapientum, Nicotianum spp., Onobrychis spp., Ornithopus spp., Oryza spp., Peltophorum africanum, Pennisetum spp., Persea gratissima, Petunia spp., Phaseolus spp., Phoenix canariensis, Phormium cookianum, Photinia spp., Picea glauca, Pinus spp., Pisum sativam, Podocarpus totara, Pogonarthria fleckii, Pogonaffhria squarrosa, Populus spp., Prosopis cineraria, Pseudotsuga menziesii, Pterolobium stellatum, Pyrus communis, Quercus spp., Rhaphiolepsis umbellata, Rhopalostylis sapida, Rhus natalensis, Ribes grossularia, Ribes spp., Robinia pseudoacacia, Rosa spp., Rubus spp., Salix spp., Schyzachyrium sanguineum, Sciadopitys vefficillata, Sequoia sempervirens, Sequoiadendron giganteum, Sorghum bicolor, Spinacia spp., Sporobolus fimbriatus, Stiburus alopecuroides, Stylosanthos humilis, Tadehagi spp, Taxodium distichum, Themeda triandra, Trifolium spp., Triticum spp., Tsuga heterophylla, Vaccinium spp., Vicia spp., Vitis vinifera, Watsonia pyramidata, Zantedeschia aethiopica, Zea mays, amaranth, artichoke, asparagus, broccoli, Brussels sprouts, cabbage, canola, carrot, cauliflower, celery, collard greens, flax, kale, lentil, oilseed rape, okra, onion, potato, rice, soybean, straw, sugar beet, sugar cane, sunflower, tomato, squash tea, maize, wheat, barely, rye, oat, peanut, pea, lentil and alfalfa, cotton, rapeseed, canola, pepper, sunflower, tobacco, eggplant, eucalyptus, a tree, an ornamental plant, a perennial grass and a forage crop. Alternatively algae and other non-Viridiplantae can be used for the methods of the present invention.
According to some embodiments of the invention, the plant used by the method of the invention is a crop plant such as rice, maize, wheat, barley, peanut, potato, sesame, olive tree, palm oil, banana, soybean, sunflower, canola, sugarcane, alfalfa, millet, leguminosae (bean, pea), flax, lupinus, rapeseed, tobacco, poplar and cotton.
Expressing the exogenous polynucleotide of the invention within the plant can be effected by transforming one or more cells of the plant with the exogenous polynucleotide, followed by generating a mature plant from the transformed cells and cultivating the mature plant under conditions suitable for expressing the exogenous polynucleotide within the mature plant.
According to some embodiments of the invention, the transformation is effected by introducing to the plant cell a nucleic acid construct which includes the exogenous polynucleotide of some embodiments of the invention and at least one promoter capable of directing transcription of the exogenous polynucleotide in the plant cell. Further details of suitable transformation approaches are provided hereinbelow.
As used herein, the term “promoter” refers to a region of DNA which lies upstream of the transcriptional initiation site of a gene to which RNA polymerase binds to initiate transcription of RNA. The promoter controls where (e.g., which portion of a plant) and/or when (e.g., at which stage or condition in the lifetime of an organism) the gene is expressed.
Any suitable promoter sequence can be used by the nucleic acid construct of the present invention. Preferably the promoter is a constitutive promoter, a tissue-specific, or an abiotic stress-inducible promoter.
Suitable constitutive promoters include, for example, CaMV 35S promoter (SEQ ID NO:2825; Odell et al., Nature 313:810-812, 1985); Arabidopsis At6669 promoter (SEQ ID NO:2823; see PCT Publication No. WO04081173A2); maize Ubi 1 (Christensen et al., Plant Sol. Biol. 18:675-689, 1992); rice actin (McElroy et al., Plant Cell 2:163-171, 1990); pEMU (Last et al., Theor. Appl. Genet. 81:581-588, 1991); CaMV 19S (Nilsson et al., Physiol. Plant 100:456-462, 1997); GOS2 (de Pater et al, Plant J November; 2(6):837-44, 1992); ubiquitin (Christensen et al, Plant Mol. Biol. 18: 675-689, 1992); Rice cyclophilin (Bucholz et al, Plant Mol Biol. 25(5):837-43, 1994); Maize H3 histone (Lepetit et al, Mol. Gen. Genet. 231: 276-285, 1992); Actin 2 (An et al, Plant J. 10(1);107-121, 1996) and Synthetic Super MAS (Ni et al., The Plant Journal 7: 661-76, 1995). Other constitutive promoters include those in U.S. Pat. Nos. 5,659,026, 5,608,149; 5,608,144; 5,604,121; 5,569,597: 5,466,785; 5,399,680; 5,268,463; and 5,608,142.
Suitable tissue-specific promoters include, but not limited to, leaf-specific promoters [such as described, for example, by Yamamoto et al., Plant J. 12:255-265, 1997; Kwon et al., Plant Physiol. 105:357-67, 1994; Yamamoto et al., Plant Cell Physiol. 35:773-778, 1994; Gotor et al., Plant J. 3:509-18, 1993; Orozco et al., Plant Mol. Biol. 23:1129-1138, 1993; and Matsuoka et al., Proc. Natl. Acad. Sci. USA 90:9586-9590, 1993], seed-preferred promoters [e.g., from seed specific genes (Simon, et al., Plant Mol. Biol. 5. 191, 1985; Scofield, et al., J. Biol. Chem. 262: 12202, 1987; Baszczynski, et al., Plant Mol. Biol. 14: 633, 1990), Brazil Nut albumin (Pearson' et al., Plant Mol. Biol. 18: 235-245, 1992), legumin (Ellis, et al. Plant Mol. Biol. 10: 203-214, 1988), Glutelin (rice) (Takaiwa, et al., Mol. Gen. Genet. 208: 15-22, 1986; Takaiwa, et al., FEBS Letts. 221: 43-47, 1987), Zein (Matzke et al Plant Mol Biol, 143). 323-32 1990), napA (Stalberg, et al, Planta 199: 515-519, 1996), Wheat SPA (Albanietal, Plant Cell, 9: 171-184, 1997), sunflower oleosin (Cummins, et al., Plant Mol. Biol. 19: 873-876, 1992)], endosperm specific promoters [e.g., wheat LMW and HMW, glutenin-1 (Mol Gen Genet 216:81-90, 1989; NAR 17:461-2), wheat a, b and g gliadins (EMBO3:1409-15, 1984), Barley ltrl promoter, barley B1, C, D hordein (Theor Appl Gen 98:1253-62, 1999; Plant J 4:343-55, 1993; Mol Gen Genet 250:750-60, 1996), Barley DOF (Mena et al, The Plant Journal, 116(1): 53-62, 1998), Biz2 (EP99106056.7), Synthetic promoter (Vicente-Carbajosa et al., Plant J. 13: 629-640, 1998), rice prolamin NRP33, rice -globulin Glb-1 (Wu et al, Plant Cell Physiology 39(8) 885-889, 1998), rice alpha-globulin REB/OHP-1 (Nakase et al. Plant Mol. Biol. 33: 513-S22, 1997), rice ADP-glucose PP (Trans Res 6:157-68, 1997), maize ESR gene family (Plant J 12:235-46, 1997), sorgum gamma-kafirin (PMB 32:1029-35, 1996)], embryo specific promoters [e.g., rice OSH1 (Sato et al, Proc. Natl. Acad. Sci. USA, 93: 8117-8122), KNOX (Postma-Haarsma of al, Plant Mol. Biol. 39:257-71, 1999), rice oleosin (Wu et at, J. Biochem., 123:386, 1998)], and flower-specific promoters [e.g., AtPRP4, chalene synthase (chsA) (Van der Meer, et al., Plant Mol. Biol. 15, 95-109, 1990), LAT52 (Twell et al Mol. Gen Genet. 217:240-245; 1989), apetala-3].
Suitable abiotic stress-inducible promoters include, but not limited to, salt-inducible promoters such as RD29A (Yamaguchi-Shinozalei et al., Mol. Gen. Genet. 236:331-340, 1993); drought-inducible promoters such as maize rab17 gene promoter (Pla et. al., Plant Mol. Biol. 21:259-266, 1993), maize rab28 gene promoter (Busk et. al., Plant J. 11:1285-1295, 1997) and maize Ivr2 gene promoter (Pelleschi et. al., Plant Mol. Biol. 39:373-380, 1999); heat-inducible promoters such as heat tomato hsp80-promoter from tomato (U.S. Pat. No. 5,187,267).
The nucleic acid construct of some embodiments of the invention can further include an appropriate selectable marker and/or an origin of replication. According to some embodiments of the invention, the nucleic acid construct utilized is a shuttle vector, which can propagate both in E. coli (wherein the construct comprises an appropriate selectable marker and origin of replication) and be compatible with propagation in cells. The construct according to the present invention can be, for example, a plasmid, a bacmid, a phagemid, a cosmid, a phage, a virus or an artificial chromosome.
The nucleic acid construct of some embodiments of the invention can be utilized to stably or transiently transform plant cells. In stable transformation, the exogenous polynucleotide is integrated into the plant genome and as such it represents a stable and inherited trait. In transient transformation, the exogenous polynucleotide is expressed by the cell transformed but it is not integrated into the genome and as such it represents a transient trait.
There are various methods of introducing foreign genes into both monocotyledonous and dicotyledonous plants (Potrykus, I., Annu. Rev. Plant. Physiol., Plant. Mol. Biol. (1991) 42:205-225; Shimamoto et al., Nature (1989) 338:274-276).
The principle methods of causing stable integration of exogenous DNA into plant genomic DNA include two main approaches:
(i) Agrobacterium-mediated gene transfer: Klee et al. (1987) Annu. Rev. Plant Physiol. 38:467-486; Klee and Rogers in Cell Culture and Somatic Cell Genetics of Plants, Vol. 6, Molecular Biology of Plant Nuclear Genes, eds. Schell, J., and Vasil, L. K., Academic Publishers, San Diego, Calif. (1989) p. 2-25; Gatenby, in Plant Biotechnology, eds. Kung, S. and Arntzen, C. J., Butterworth Publishers, Boston, Mass. (1989) p. 93-112.
(ii) Direct DNA uptake: Paszkowski et al., in Cell Culture and Somatic Cell Genetics of Plants, Vol. 6, Molecular Biology of Plant Nuclear Genes eds. Schell, J., and Vasil, L. K., Academic Publishers, San Diego, Calif. (1989) p. 52-68; including methods for direct uptake of DNA into protoplasts, Toriyama, K. et al. (1988) Bio/Technology 6:1072-1074. DNA uptake induced by brief electric shock of plant cells: Zhang et al. Plant Cell Rep. (1988) 7:379-384. Fromm et al. Nature (1986) 319:791-793. DNA injection into plant cells or tissues by particle bombardment, Klein et al. Bio/Technology (1988) 6:559-563; McCabe et al. Bio/Technology (1988) 6:923-926; Sanford, Physiol. Plant. (1990) 79:206-209; by the use of micropipette systems: Neuhaus et al., Theor. Appl. Genet. (1987) 75:30-36; Neuhaus and Spangenberg, Physiol. Plant. (1990) 79:213-217; glass fibers or silicon carbide whisker transformation of cell cultures, embryos or callus tissue, U.S. Pat. No. 5,464,765 or by the direct incubation of DNA with germinating pollen, DeWet et al. in Experimental Manipulation of Ovule Tissue, eds. Chapman, G. P. and Mantell, S. H. and Daniels, W. Longman, London, (1985) p. 197-209; and Ohta, Proc. Natl. Acad. Sci. USA (1986) 83:715-719.
The Agrobacterium system includes the use of plasmid vectors that contain defined DNA segments that integrate into the plant genomic DNA. Methods of inoculation of the plant tissue vary depending upon the plant species and the Agrobacterium delivery system. A widely used approach is the leaf disc procedure which can be performed with any tissue explant that provides a good source for initiation of whole plant differentiation. See, e.g., Horsch et al. in Plant Molecular Biology Manual A5, Kluwer Academic Publishers, Dordrecht (1988) p. 1-9. A supplementary approach employs the Agrobacterium delivery system in combination with vacuum infiltration. The Agrobacterium system is especially viable in the creation of transgenic dicotyledonous plants.
There are various methods of direct DNA transfer into plant cells. In electroporation, the protoplasts are briefly exposed to a strong electric field. In microinjection, the DNA is mechanically injected directly into the cells using very small micropipettes. In microparticle bombardment, the DNA is adsorbed on microprojectiles such as magnesium sulfate crystals or tungsten particles, and the microprojectiles are physically accelerated into cells or plant tissues.
Following stable transformation plant propagation is exercised. The most common method of plant propagation is by seed. Regeneration by seed propagation, however, has the deficiency that due to heterozygosity there is a lack of uniformity in the crop, since seeds are produced by plants according to the genetic variances governed by Mendelian rules. Basically, each seed is genetically different and each will grow with its own specific traits. Therefore, it is preferred that the transformed plant be produced such that the regenerated plant has the identical traits and characteristics of the parent transgenic plant. Therefore, it is preferred that the transformed plant be regenerated by micropropagation which provides a rapid, consistent reproduction of the transformed plants.
Micropropagation is a process of growing new generation plants from a single piece of tissue that has been excised from a selected parent plant or cultivar. This process permits the mass reproduction of plants having the preferred tissue expressing the fusion protein. The new generation plants which are produced are genetically identical to, and have all of the characteristics of, the original plant. Micropropagation allows mass production of quality plant material in a short period of time and offers a rapid multiplication of selected cultivars in the preservation of the characteristics of the original transgenic or transformed plant. The advantages of cloning plants are the speed of plant multiplication and the quality and uniformity of plants produced.
Micropropagation is a multi-stage procedure that requires alteration of culture medium or growth conditions between stages. Thus, the micropropagation process involves four basic stages: Stage one, initial tissue culturing; stage two, tissue culture multiplication; stage three, differentiation and plant formation; and stage four, greenhouse culturing and hardening. During stage one, initial tissue culturing, the tissue culture is established and certified contaminant-free. During stage two, the initial tissue culture is multiplied until a sufficient number of tissue samples are produced to meet production goals. During stage three, the tissue samples grown in stage two are divided and grown into individual plantlets. At stage four, the transformed plantlets are transferred to a greenhouse for hardening where the plants' tolerance to light is gradually increased so that it can be grown in the natural environment.
According to some embodiments of the invention, the transgenic plants are generated by transient transformation of leaf cells, meristematic cells or the whole plant.
Transient transformation can be effected by any of the direct DNA transfer methods described above or by viral infection using modified plant viruses.
Viruses that have been shown to be useful for the transformation of plant hosts include CaMV, Tobacco mosaic virus (TMV), brome mosaic virus (BMV) and Bean Common Mosaic Virus (BV or BCMV). Transformation of plants using plant viruses is described in U.S. Pat. No. 4,855,237 (bean golden mosaic virus; BGV), EP-A 67,553 (TMV), Japanese Published Application No. 63-14693 (TMV), EPA 194,809 (BV), EPA 278,667 (BV); and Gluzman, Y. et al., Communications in Molecular Biology: Viral Vectors, Cold Spring Harbor Laboratory, New York, pp. 172-189 (1988). Pseudovirus particles for use in expressing foreign DNA in many hosts, including plants are described in WO 87/06261.
According to some embodiments of the invention, the virus used for transient transformations is avirulent and thus is incapable of causing severe symptoms such as reduced growth rate, mosaic, ring spots, leaf roll, yellowing, streaking, pox formation, tumor formation and pitting. A suitable avirulent virus may be a naturally occurring avirulent virus or an artificially attenuated virus. Virus attenuation may be effected by using methods well known in the art including, but not limited to, sub-lethal heating, chemical treatment or by directed mutagenesis techniques such as described, for example, by Kurihara and Watanabe (Molecular Plant Pathology 4:259-269, 2003), Gal-on et al. (1992), Atreya et al. (1992) and Huet et al. (1994).
Suitable virus strains can be obtained from available sources such as, for example, the American Type culture Collection (ATCC) or by isolation from infected plants. Isolation of viruses from infected plant tissues can be effected by techniques well known in the art such as described, for example by Foster and Tatlor, Eds. “Plant Virology Protocols: From Virus Isolation to Transgenic Resistance (Methods in Molecular Biology (Humana Pr), Vol 81)”, Humana Press, 1998. Briefly, tissues of an infected plant believed to contain a high concentration of a suitable virus, preferably young leaves and flower petals, are ground in a buffer solution (e.g., phosphate buffer solution) to produce a virus infected sap which can be used in subsequent inoculations.
Construction of plant RNA viruses for the introduction and expression of non-viral exogenous polynucleotide sequences in plants is demonstrated by the above references as well as by Dawson, W. O. et al., Virology (1989) 172:285-292; Takamatsu et al. EMBO J. (1987) 6:307-311; French et al. Science (1986) 231:1294-1297; Takamatsu et al. FEBS Letters (1990) 269:73-76; and U.S. Pat. No. 5,316,931.
When the virus is a DNA virus, suitable modifications can be made to the virus itself. Alternatively, the virus can first be cloned into a bacterial plasmid for ease of constructing the desired viral vector with the foreign DNA. The virus can then be excised from the plasmid. If the virus is a DNA virus, a bacterial origin of replication can be attached to the viral DNA, which is then replicated by the bacteria. Transcription and translation of this DNA will produce the coat protein which will encapsidate the viral DNA. If the virus is an RNA virus, the virus is generally cloned as a cDNA and inserted into a plasmid. The plasmid is then used to make all of the constructions. The RNA virus is then produced by transcribing the viral sequence of the plasmid and translation of the viral genes to produce the coat protein(s) which encapsidate the viral RNA.
In one embodiment, a plant viral polynucleotide is provided in which the native coat protein coding sequence has been deleted from a viral polynucleotide, a non-native plant viral coat protein coding sequence and a non-native promoter, preferably the subgenomic promoter of the non-native coat protein coding sequence, capable of expression in the plant host, packaging of the recombinant plant viral polynucleotide, and ensuring a systemic infection of the host by the recombinant plant viral polynucleotide, has been inserted. Alternatively, the coat protein gene may be inactivated by insertion of the non-native polynucleotide sequence within it, such that a protein is produced. The recombinant plant viral polynucleotide may contain one or more additional non-native subgenomic promoters. Each non-native subgenomic promoter is capable of transcribing or expressing adjacent genes or polynucleotide sequences in the plant host and incapable of recombination with each other and with native subgenomic promoters. Non-native (foreign) polynucleotide sequences may be inserted adjacent the native plant viral subgenomic promoter or the native and a non-native plant viral subgenomic promoters if more than one polynucleotide sequence is included. The non-native polynucleotide sequences are transcribed or expressed in the host plant under control of the subgenomic promoter to produce the desired products.
In a second embodiment, a recombinant plant viral polynucleotide is provided as in the first embodiment except that the native coat protein coding sequence is placed adjacent one of the non-native coat protein subgenomic promoters instead of a non-native coat protein coding sequence.
In a third embodiment, a recombinant plant viral polynucleotide is provided in which the native coat protein gene is adjacent its subgenomic promoter and one or more non-native subgenomic promoters have been inserted into the viral polynucleotide. The inserted non-native subgenomic promoters are capable of transcribing or expressing adjacent genes in a plant host and are incapable of recombination with each other and with native subgenomic promoters. Non-native polynucleotide sequences may be inserted adjacent the non-native subgenomic plant viral promoters such that the sequences are transcribed or expressed in the host plant under control of the subgenomic promoters to produce the desired product.
In a fourth embodiment, a recombinant plant viral polynucleotide is provided as in the third embodiment except that the native coat protein coding sequence is replaced by a non-native coat protein coding sequence.
The viral vectors are encapsidated by the coat proteins encoded by the recombinant plant viral polynucleotide to produce a recombinant plant virus. The recombinant plant viral polynucleotide or recombinant plant virus is used to infect appropriate host plants. The recombinant plant viral polynucleotide is capable of replication in the host, systemic spread in the host, and transcription or expression of foreign gene(s) (exogenous polynucleotide) in the host to produce the desired protein.
Techniques for inoculation of viruses to plants may be found in Foster and Taylor, eds. “Plant Virology Protocols: From Virus Isolation to Transgenic Resistance (Methods in Molecular Biology (Humana Pr), Vol 81)”, Humana Press, 1998; Maramorosh and Koprowski, eds. “Methods in Virology” 7 vols, Academic Press, New York 1967-1984; Hill, S. A. “Methods in Plant Virology”, Blackwell, Oxford, 1984; Walkey, D. G. A. “Applied Plant Virology”, Wiley, New York, 1985; and Kado and Agrawa, eds. “Principles and Techniques in Plant Virology”, Van Nostrand-Reinhold, New York.
In addition to the above, the polynucleotide of the present invention can also be introduced into a chloroplast genome thereby enabling chloroplast expression.
A technique for introducing exogenous polynucleotide sequences to the genome of the chloroplasts is known. This technique involves the following procedures. First, plant cells are chemically treated so as to reduce the number of chloroplasts per cell to about one. Then, the exogenous polynucleotide is introduced via particle bombardment into the cells with the aim of introducing at least one exogenous polynucleotide molecule into the chloroplasts. The exogenous polynucleotides selected such that it is integratable into the chloroplast's genome via homologous recombination which is readily effected by enzymes inherent to the chloroplast. To this end, the exogenous polynucleotide includes, in addition to a gene of interest, at least one polynucleotide stretch which is derived from the chloroplast's genome. In addition, the exogenous polynucleotide includes a selectable marker, which serves by sequential selection procedures to ascertain that all or substantially all of the copies of the chloroplast genomes following such selection will include the exogenous polynucleotide. Further details relating to this technique are found in U.S. Pat. Nos. 4,945,050; and 5,693,507 which are incorporated herein by reference. A polypeptide can thus be produced by the protein expression system of the chloroplast and become integrated into the chloroplast's inner membrane.
Since abiotic stress tolerance, water use efficiency, fertilizer use efficiency, growth, biomass, yield and/or vigor in plants can involve multiple genes acting additively or in synergy (see, for example, in Quesda et al., Plant Physiol. 130:951-063, 2002), the present invention also envisages expressing a plurality of exogenous polynucleotides in a single host plant to thereby achieve superior effect on abiotic stress tolerance, water use efficiency, fertilizer use efficiency, growth, biomass, yield and/or vigor.
Expressing a plurality of exogenous polynucleotides in a single host plant can be effected by co-introducing multiple nucleic acid constructs, each including a different exogenous polynucleotide, into a single plant cell. The transformed cell can than be regenerated into a mature plant using the methods described hereinabove.
Alternatively, expressing a plurality of exogenous polynucleotides in a single host plant can be effected by co-introducing into a single plant-cell a single nucleic-acid construct including a plurality of different exogenous polynucleotides. Such a construct can be designed with a single promoter sequence which can transcribe a polycistronic messenger RNA including all the different exogenous polynucleotide sequences. To enable co-translation of the different polypeptides encoded by the polycistronic messenger RNA, the polynucleotide sequences can be inter-linked via an internal ribosome entry site (IRES) sequence which facilitates translation of polynucleotide sequences positioned downstream of the IRES sequence. In this case, a transcribed polycistronic RNA molecule encoding the different polypeptides described above will be translated from both the capped 5′ end and the two internal IRES sequences of the polycistronic RNA molecule to thereby produce in the cell all different polypeptides. Alternatively, the construct can include several promoter sequences each linked to a different exogenous polynucleotide sequence.
The plant cell transformed with the construct including a plurality of different exogenous polynucleotides, can be regenerated into a mature plant, using the methods described hereinabove.
Alternatively, expressing a plurality of exogenous polynucleotides in a single host plant can be effected by introducing different nucleic acid constructs, including different exogenous polynucleotides, into a plurality of plants. The regenerated transformed plants can then be cross-bred and resultant progeny selected for superior abiotic stress tolerance, water use efficiency, fertilizer use efficiency, growth, biomass, yield and/or vigor traits, using conventional plant breeding techniques.
Thus, the invention encompasses plants exogenously expressing (as described above) the polynucleotide(s) and/or polypeptide(s) of the invention. Once expressed within the plant cell or the entire plant, the level of the polypeptide encoded by the exogenous polynucleotide can be determined by methods well known in the art such as, activity assays, Western blots using antibodies capable of specifically binding the polypeptide, Enzyme-Linked ImmunoSorbent Assay (ELISA), radio-immuno-assays (RIA), immunohistochemistry, immunocytochemistry, immunofluorescence and the like.
Methods of determining the level in the plant of the RNA transcribed from the exogenous polynucleotide are well known in the art and include, for example, Northern blot analysis, reverse transcription polymerase chain reaction (RT-PCR) analysis (including quantitative, semi-quantitative or real-time RT-PCR) and RNA-in situ hybridization.
As mentioned, the polypeptide according to some embodiments of the invention, functions as a water channel. Thus, the invention according to some embodiments encompasses functional equivalents of the polypeptide (e.g., polypeptides capable of the biological activity of a water channel) which can be identified by functional assays (e.g., being capable of transporting water in a plant) using e.g., a cell-swelling assay (Meng, Q. X. et al. 2008. Cell Physiol Biochem, 21. pp. 123-128).
The polynucleotides and polypeptides described hereinabove can be used in a wide range of economical plants, in a safe and cost effective manner.
The effect of the transgene (the exogenous polynucleotide encoding the polypeptide) on abiotic stress tolerance, water use efficiency, fertilizer use efficiency, growth, biomass, yield and/or vigor can be determined using known methods.
Abiotic stress tolerance—Transformed (i.e., expressing the transgene) and non-transformed (wild type) plants are exposed to an abiotic stress condition, such as water deprivation, suboptimal temperature (low temperature, high temperature), nutrient deficiency, nutrient excess, a salt stress condition, osmotic stress, heavy metal toxicity, anaerobiosis, atmospheric pollution and UV irradiation.
Salinity tolerance assay—Transgenic plants with tolerance to high salt concentrations are expected to exhibit better germination, seedling vigor or growth in high salt. Salt stress can be effected in many ways such as, for example, by irrigating the plants with a hyperosmotic solution, by cultivating the plants hydroponically in a hyperosmotic growth solution (e.g., Hoagland solution), or by culturing the plants in a hyperosmotic growth medium [e.g., 50% Murashige-Skoog medium (MS medium)]. Since different plants vary considerably in their tolerance to salinity, the salt concentration in the irrigation water, growth solution, or growth medium can be adjusted according to the specific characteristics of the specific plant cultivar or variety, so as to inflict a mild or moderate effect on the physiology and/or morphology of the plants (for guidelines as to appropriate concentration see, Bernstein and Kafkafi, Root Growth Under Salinity Stress In: Plant Roots, The Hidden Half 3rd ed. Waisel Y, Eshel A and Kafkafi U. (editors) Marcel Dekker Inc., New York, 2002, and reference therein).
For example, a salinity tolerance test can be performed by irrigating plants at different developmental stages with increasing concentrations of sodium chloride (for example 50 mM, 100 mM, 200 mM, 400 mM NaCl) applied from the bottom and from above to ensure even dispersal of salt. Following exposure to the stress condition the plants are frequently monitored until substantial physiological and/or morphological effects appear in wild type plants. Thus, the external phenotypic appearance, degree of wilting and overall success to reach maturity and yield progeny are compared between control and transgenic plants. Quantitative parameters of tolerance measured include, but are not limited to, the average wet and dry weight, the weight of the seeds yielded, the average seed size and the number of seeds produced per plant. Transformed plants not exhibiting substantial physiological and/or morphological effects, or exhibiting higher biomass than wild-type plants, are identified as abiotic stress tolerant plants.
Osmotic tolerance test—Osmotic stress assays (including sodium chloride and mannitol assays) are conducted to determine if an osmotic stress phenotype was sodium chloride-specific or if it was a general osmotic stress related phenotype. Plants which are tolerant to osmotic stress may have more tolerance to drought and/or freezing. For salt and osmotic stress germination experiments, the medium is supplemented for example with 50 mM, 100 mM, 200 mM NaCl or 100 mM, 200 mM NaCl, 400 mM mannitol. See also Example 5 of the Examples section which follows.
Drought tolerance assay/Osmoticum assay—Tolerance to drought is performed to identify the genes conferring better plant survival after acute water deprivation. To analyze whether the transgenic plants are more tolerant to drought, an osmotic stress produced by the non-ionic osmolyte sorbitol in the medium can be performed. Control and transgenic plants are germinated and grown in plant-agar plates for 4 days, after which they are transferred to plates containing 500 mM sorbitol. The treatment causes growth retardation, then both control and transgenic plants are compared, by measuring plant weight (wet and dry), yield, and by growth rates measured as time to flowering.
Conversely, soil-based drought screens are performed with plants overexpressing the polynucleotides detailed above. Seeds from control Arabidopsis plants, or other transgenic plants overexpressing the polypeptide of the invention are germinated and transferred to pots. Drought stress is obtained after irrigation is ceased accompanied by placing the pots on absorbent paper to enhance the soil-drying rate. Transgenic and control plants are compared to each other when the majority of the control plants develop severe wilting. Plants are re-watered after obtaining a significant fraction of the control plants displaying a severe wilting. Plants are ranked comparing to controls for each of two criteria: tolerance to the drought conditions and recovery (survival) following re-watering.
Cold stress tolerance—To analyze cold stress, mature (25 day old) plants are transferred to 4° C. chambers for 1 or 2 weeks, with constitutive light. Later on plants are moved back to greenhouse. Two weeks later damages from chilling period, resulting in growth retardation and other phenotypes, are compared between both control and transgenic plants, by measuring plant weight (wet and dry), and by comparing growth rates measured as time to flowering, plant size, yield, and the like.
Heat stress tolerance—Heat stress tolerance is achieved by exposing the plants to temperatures above 34° C. for a certain period. Plant tolerance is examined after transferring the plants back to 22° C. for recovery and evaluation after 5 days relative to internal controls (non-transgenic plants) or plants not exposed to neither cold or heat stress.
Germination tests—Germination tests compare the percentage of seeds from transgenic plants that could complete the germination process to the percentage of seeds from control plants that are treated in the same manner. Normal conditions are considered for example, incubations at 22° C. under 22-hour light 2-hour dark daily cycles. Evaluation of germination and seedling vigor is conducted between 4 and 14 days after planting. The basal media is 50% MS medium (Murashige and Skoog, 1962 Plant Physiology 15, 473-497).
Germination is checked also at unfavorable conditions such as cold (incubating at temperatures lower than 10° C. instead of 22° C.) or using seed inhibition solutions that contain high concentrations of an osmolyte such as sorbitol (at concentrations of 50 mM, 100 mM, 200 mM, 300 mM, 500 mM, and up to 1000 mM) or applying increasing concentrations of salt (of 50 mM, 100 mM, 200 mM, 300 mM, 500 mM NaCl).
Water use efficiency—can be determined as the biomass produced per unit transpiration. To analyze WUE, leaf relative water content can be measured in control and transgenic plants. Fresh weight (FW) is immediately recorded; then leaves are soaked for 8 hours in distilled water at room temperature in the dark, and the turgid weight (TW) is recorded. Total dry weight (DW) is recorded after drying the leaves at 60° C. to a constant weight. Relative water content (RWC) is calculated according to the following Formula I:
(FW−DW/TW'DW)×100 Formula I
Fertilizer use efficiency—To analyze whether the transgenic plants are more responsive to fertilizers, plants are grown in agar plates or pots with a limited amount of fertilizer, as described, for example, in Example 6, hereinbelow and in Yanagisawa et al (Proc Natl Acad Sci USA. 2004; 101:7833-8). The plants are analyzed for their overall size, time to flowering, yield, protein content of shoot and/or grain. The parameters checked are the overall size of the mature plant, its wet and dry weight, the weight of the seeds yielded, the average seed size and the number of seeds produced per plant. Other parameters that may be tested are: the chlorophyll content of leaves (as nitrogen plant status and the degree of leaf verdure is highly correlated), amino acid and the total protein content of the seeds or other plant parts such as leaves or shoots, oil content, etc. Similarly, instead of providing nitrogen at limiting amounts, phosphate or potassium can be added at increasing concentrations. Again, the same parameters measured are the same as listed above. In this way, nitrogen use efficiency (NUE), phosphate use efficiency (PUE) and potassium use efficiency (KUE) are assessed, checking the ability of the transgenic plants to thrive under nutrient restraining conditions.
Nitrogen determination—The procedure for N (nitrogen) concentration determination in the structural parts of the plants involves the potassium persulfate digestion method to convert organic N to NO₃ ⁻ (Purcell and King 1996 Argon. J. 88:111-113, the modified Cd⁻ mediated reduction of NO₃ ⁻ to NO2⁻ (Vodovotz 1996 Biotechniques 20:390-394) and the measurement of nitrite by the Griess assay (Vodovotz 1996, supra). The absorbance values are measured at 550 nm against a standard curve of NaNO₂. The procedure is described in details in Samonte et al. 2006 Agron. J. 98:168-176.
Grain protein concentration—Grain protein content (g grain protein m⁻²) is estimated as the product of the mass of grain N (g grain N m⁻²) multiplied by the N/protein conversion ratio of k-5.13 (Mosse 1990, supra). The grain protein concentration is estimated as the ratio of grain protein content per unit mass of the grain (g grain protein kg⁻¹grain).
Oil content—The oil content of a plant can be determined by extraction of the oil from the seed or the vegetative portion of the plant. Briefly, lipids (oil) can be removed from the plant (e.g., seed) by grinding the plant tissue in the presence of specific solvents (e.g., hexane or petroleum ether) and extracting the oil in a continuous extractor. Indirect oil content analysis can be carried out using various known methods such as Nuclear Magnetic Resonance (NMR) Spectroscopy, which measures the resonance energy absorbed by hydrogen atoms in the liquid state of the sample [See for example, Conway T F. and Earle F R., 1963, Journal of the American Oil Chemists' Society; Springer Berlin/Heidelberg, ISSN: 0003-021X (Print) 1558-9331 (Online)]; the Near Infrared (NI) Spectroscopy, which utilizes the absorption of near infrared energy (1100-2500 nm) by the sample; and a method described in WO/2001/023884, which is based on extracting oil a solvent, evaporating the solvent in a gas stream which forms oil particles, and directing a light into the gas stream and oil particles which forms a detectable reflected light.
The plant vigor can be calculated by the increase in growth parameters such as leaf area, fiber length, rosette diameter, plant fresh weight and the like per time.
The growth rate can be measured using digital analysis of growing plants. For example, images of plants growing in greenhouse on plot basis can be captured every 3 days and the rosette area can be calculated by digital analysis. Rosette area growth is calculated using the difference of rosette area between days of sampling divided by the difference in days between samples.
Measurements of seed yield can be done by collecting the total seeds from 8-16 plants together, weighting them using analytical balance and dividing the total weight by the number of plants. Seed per growing area can be calculated in the same manner while taking into account the growing area given to a single plant. Increase seed yield per growing area could be achieved by increasing seed yield per plant, and/or by increasing number of plants capable of growing in a given area.
Evaluation of the seed yield per plant can be done by measuring the amount (weight or size) or quantity (i.e., number) of dry seeds produced and harvested from 8-16 plants and divided by the number of plants.
Evaluation of growth rate can be done by measuring plant biomass produced, rosette area, leaf size or root length per time (can be measured in cm²per day of leaf area).
Fiber length can be measured using fibrograph. The fibrograph system was used to compute length in terms of “Upper Half Mean” length. The upper half mean (UHM) is the average length of longer half of the fiber distribution. The fibrograph measures length in span lengths at a given percentage point (Hypertext Transfer Protocol://World Wide Web (dot) cottoninc (dot) com/ClassificationofCotton/?Pg=4#Length).
Thus, the present invention is of high agricultural value for promoting the yield of commercially desired crops (e.g., biomass of vegetative organ such as poplar wood, or reproductive organ such as number of seeds or seed biomass).
As used herein the term “about” refers to ±10%.
The terms “comprises”, “comprising”, “includes”, “including”, “having” and their conjugates mean “including but not limited to”.
The term “consisting of means “including and limited to”.
The term “consisting essentially of” means that the composition, method or structure may include additional ingredients, steps and/or parts, but only if the additional ingredients, steps and/or parts do not materially alter the basic and novel characteristics of the claimed composition, method or structure.
As used herein, the singular form “a”, “an” and “the” include plural references unless the context clearly dictates otherwise. For example, the term “a compound” or “at least one compound” may include a plurality of compounds, including mixtures thereof.
Throughout this application, various embodiments of this invention may be presented in a range format. It should be understood that the description in range format is merely for convenience and brevity and should not be construed as an inflexible limitation on the scope of the invention. Accordingly, the description of a range should be considered to have specifically disclosed all the possible subranges as well as individual numerical values within that range. For example, description of a range such as from 1 to 6 should be considered to have specifically disclosed subranges such as from 1 to 3, from 1 to 4, from 1 to 5, from 2 to 4, from 2 to 6, from 3 to 6 etc., as well as individual numbers within that range, for example, 1, 2, 3, 4, 5, and 6. This applies regardless of the breadth of the range.
Whenever a numerical range is indicated herein, it is meant to include any cited numeral (fractional or integral) within the indicated range. The phrases “ranging/ranges between” a first indicate number and a second indicate number and “ranging/ranges from” a first indicate number “to” a second indicate number are used herein interchangeably and are meant to include the first and second indicated numbers and all the fractional and integral numerals therebetween.
As used herein the term “method” refers to manners, means, techniques and procedures for accomplishing a given task including, but not limited to, those manners, means, techniques and procedures either known to, or readily developed from known manners, means, techniques and procedures by practitioners of the chemical, pharmacological, biological, biochemical and medical arts.
It is appreciated that certain features of the invention, which are, for clarity, described in the context of separate embodiments, may also be provided in combination in a single embodiment. Conversely, various features of the invention, which are, for brevity, described in the context of a single embodiment, may also be provided separately or in any suitable subcombination or as suitable in any other described embodiment of the invention. Certain features described in the context of various embodiments are not to be considered essential features of those embodiments, unless the embodiment is inoperative without those elements.
Various embodiments and aspects of the present invention as delineated hereinabove and as claimed in the claims section below find experimental support in the following examples.

EXAMPLES

Reference is now made to the following examples, which together with the above descriptions illustrate some embodiments of the invention in a non limiting fashion.
Generally, the nomenclature used herein and the laboratory procedures utilized in the present invention include molecular, biochemical, microbiological and recombinant DNA techniques. Such techniques are thoroughly explained in the literature. See, for example, “Molecular Cloning: A laboratory Manual” Sambrook et al., (1989); “Current Protocols in Molecular Biology” Volumes I-III Ausubel, R. M., ed. (1994); Ausubel et al., “Current Protocols in Molecular Biology”, John Wiley and Sons, Baltimore, Md. (1989); Perbal, “A Practical Guide to Molecular Cloning”, John Wiley & Sons, New York (1988); Watson et al., “Recombinant DNA”, Scientific American Books, New York; Birren et al. (eds) “Genome Analysis: A Laboratory Manual Series”, Vols. 1-4, Cold Spring Harbor Laboratory Press, New York (1998); methodologies as set forth in U.S. Pat. Nos. 4,666,828; 4,683,202; 4,801,531; 5,192,659 and 5,272,057; “Cell Biology: A Laboratory Handbook”, Volumes I-III Cellis, J. E., ed. (1994); “Current Protocols in Immunology” Volumes I-III Coligan J. E., ed. (1994); Stites et al. (eds), “Basic and Clinical Immunology” (8th Edition), Appleton & Lange, Norwalk, Conn. (1994); Mishell and Shiigi (eds), “Selected Methods in Cellular Immunology”, W. H. Freeman and Co., New York (1980); available immunoassays are extensively described in the patent and scientific literature, see, for example, U.S. Pat. Nos. 3,791,932; 3,839,153; 3,850,752; 3,850,578; 3,853,987; 3,867,517; 3,879,262; 3,901,654; 3,935,074; 3,984,533; 3,996,345; 4,034,074; 4,098,876; 4,879,219; 5,011,771 and 5,281,521; “Oligonucleotide Synthesis” Gait, M. J., ed. (1984); “Nucleic Acid Hybridization” Hames, B. D., and Higgins S. J., eds. (1985); “Transcription and Translation” Hames, B. D., and Higgins S. J., Eds. (1984); “Animal Cell Culture” Freshney, R. I., ed. (1986); “Immobilized Cells and Enzymes” IRL Press, (1986); “A Practical Guide to Molecular Cloning” Perbal, B., (1984) and “Methods in Enzymology” Vol. 1-317, Academic Press; “PCR Protocols: A Guide To Methods And Applications”, Academic Press, San Diego, Calif. (1990); Marshak et al., “Strategies for Protein Purification and Characterization—A Laboratory Course Manual” CSHL Press (1996); all of which are incorporated by reference as if fully set forth herein. Other general references are provided throughout this document. The procedures therein are believed to be well known in the art and are provided for the convenience of the reader. All the information contained therein is incorporated herein by reference.

Example 1

Identification of AQP Genes using Digital Expression Analysis and Cross-Species Comparative Genomic

The large number of AQPs in plants and the contradictory results obtained when AQPs were overexpressed in plants demonstrate the need to selectively identify the AQP genes which can improve water use efficiency (WUE) in plants, lead to increased yield and biomass under abiotic stress as well as under favorable conditions.
Under unfavorable stress conditions, some biological activities of the plant are stopped or reduced, while others, not earlier active, initiate. Still, some of the activities, which are vital for plant survival, are maintained. One hypothesis is that key genes needed for plants to maintain vital activities under unfavorable conditions would be active under broad spectrum of biotic and abiotic stresses.
To test this hypothesis and to identify the key AQP genes having the potential to improve plant performance under different biotic and/or abiotic stress conditions (e.g., salt or drought stress) a combination of digital expression analysis (also known as Electronic Northern blot) and cross-species comparative genomics was performed. The database used was available from NCBI (Hypertext Transfer Protocol://World Wide Web (dot) ncbi (dot) nlm (dot) nih (dot) gov/dbEST/) and included 7.2 million expressed sequence tags (ESTs) from 1,195 relevant EST's libraries originated from 15 different species, including both monocot and dicot species, namely: Arabidopsis, barley, Brassica rapa, cotton, grape, maize, medicago, poplar, potato, rice, sorghum, soybean, sugarcane, tomato and wheat.
Tomato plants were selected as a model plant based on the high quality tomato database from several tomato species which can be used for data-mining and the present inventors' experience in using the tomato genome as a model plant. In addition, the relatively high salt tolerance exhibited by various tomato species makes the tomato genome an excellent candidate for identifying new stress tolerance mechanisms. Moreover, tomato is not only used as a model plant for genetic studies, it is also used as an important crop with well-defined yield parameters, which can be used to distinguish between genes affecting abiotic-stress tolerance and genes preventing yield loss under abiotic-stress conditions.
Gene analysis and data mining—For gene analysis and data mining the bioinformatic filtering approach used had three phases:
1. Clustering and assembly: EST and mRNA sequences of each of the 15 species were extracted from GenBank versions 157, 160, 161, 162, 164, 165, 166, clustered and assembled using Compugen's LEADS clustering and assembly platform (Compugen Ltd., Tel Aviv, Israel; Yelin et. al. 2003, Nature Biotechnology 21, 379-85). Automatically extracted EST library annotations were manually accurated and classified by anatomy, developmental stage, abiotic/biotic stress treatment and cultivars. The results were loaded into Oracle database. The predicted proteins were then annotated using InterPro(2) (Hypertext Transfer Protocol://World Wide Web (dot) ebi (dot) ac (dot) uk/interpro/).
2. Selection of clusters—All clusters that contained the Major intrinsic protein domain (IPR000425) were selected for further analysis (n=1,114).
3. Obtaining expression profile of the clusters—By digital expression approach the expression profile of all clusters was obtained in terms of plant anatomy (i.e., in what tissues/organs the gene was expressed), developmental stage (i.e., the developmental stages at which a gene can be found) and profile of treatment (provides the physiological conditions under which a gene is expressed such as drought, cold, pathogen infection, etc).
Digital expression computations was calculated as follows: over-expression fold was computed as m/(n*M/N), where “N” is total number of ESTs of specific organism; “M is number of ESTs in a given library/tissue/category; “n” is total number of ESTs in a given contig; “m” is the number of ESTs from the library/tissue/category in the contig; P-value was computed using Exact Fisher Test statistic. The combined P-value for over-expression in both Root and Abiotic stresses conditions was computed as 1−(1-p1)×(1-p2). 1,114 different AQP genes were identified in the inter species transcriptional databases. For the data mining process, the present inventors used a combination of two approaches: selection of AQP clusters showing significant over expression (EST distribution versus normal is more than two folds; statistical significance of over-expression−p Value<0.05) either in roots compared to shoots or under various abiotic stresses (including drought, cold, salinity, heat, chemical treatments, etc.), compared to non stress control. It was found that ESTs of about 9% of the AQP genes were significantly overrepresented in roots and 3.5% of them were induced under different abiotic stresses. AQP genes which are highly overrepresented in roots were selected since plants with an efficient root system are expected to capture more water from a drying soil. In addition, AQP genes which are overrepresented in various abiotic stresses such as nutrient deficiency, heat, salinity and heavy metal stresses and biotic stresses such as application of elicitors and pathogens were selected considering that they can provide high tolerance to a wide spectrum of stresses.
The same set of 1,114 AQPs was classified according to the accepted groups known in the literature: first into the four major sub-groups: PIPs, TIPs, NIPs and SIPs, and a second classification divided these four sub-groups into eleven sub-groups according to their homology in amino acid sequences. A Fisher's exact test was then used to identify subgroups having significant EST over-presentation both in roots and upon exposure to different abiotic stresses. As shown in Table 1, hereinbelow, from the eleven subgroups, only the TIP2 subgroup showed a significant EST overrepresentation both in roots and upon exposure to abiotic stresses (P-value 1.7×10⁻⁵and 1.6×10⁻³, respectively).

TABLE 1

AQP type distribution and over-expression in roots and abiotic stresses

		Exposure to
	Roots	abiotic stresses

	Total No. of	No. of over-			No. of over-
AQP	genes in	expressed	% over-	P-	expressed	% over-	P-
type	database	genes	expressed/all	value	genes	expressed/all	value

PIP1	243	26	10.7	0.13	10	4.1	0.34
PIP2	336	25	7.4	0.87	12	3.6	0.53
PIP3	11	0	0.0	1	0	0	1
SIP1	39	0	0.0	1	0	0	1
SIP2	16	0	0.0	1	0	0	1
TIP1	152	11	7.2	0.8	3	2	0.92
TIP2	101	22	21.8	1.70E−05	10	9.9	1.6E−0.3
TIP3	29	0	0.0	1	0	0	1
TIP4	48	5	10.4	0.41	1	2.1	0.83
TIP5	3	0	0.0	1	0	0	1
NIP	136	8	5.9	0.93	3	2.2	0.88
Total	1114	97			39

Table 1.

These results suggest that over-expression and/or protein over-accumulation of the Tip2 subgroup can improve plant water use efficiency, ABST and yield.
Genes of the Tip2 subgroup are highly expressed in roots and in abiotic stresses—As shown in Table 1, hereinabove, the TIP2 subgroup (or subfamily) is highly expressed in roots and in abiotic stresses. The TIP2 subgroup is found in 38 plant species and other organisms (nucleic acid SEQ ID NOs: 1, 2, 19, 20-22; Table 2), available in public databases [Hypertext Transfer Protocol://World Wide Web (dot) ncbi (dot) nlm (dot) nih (dot) gov/dbEST/]. In tomato, the TIP2 gene was highly expressed in roots (6 fold, p≦1.01 E-24) and in both biotic (2 fold, p≦4.6 E-02) and abiotic stresses (4.5 fold, p≦E-02) (data not shown).
Identification of a short consensus sequence of the Tip2 sub family—While comparing the consensus amino-acid sequences of Aquaporins, a short consensus sequence was identified which is unique to proteins of the Tip2 sub-family. The present inventors have suggested that this motif has an important role in managing water use efficiency (WUE), and when over-expressed in a plant can confer ABST and improved yield. The amino-acid consensus sequence identified is TLXFXFAGVGS (SEQ ID NO:2826), wherein X stands for any amino acid.
In addition, other genes of the aquaporin gene family were identified by bioinformatics tools as improving ABST and yield, based on combined digital gene expression profile in roots, tissues with low water levels (such as seed and pollen) and under abiotic stress conditions. These include SEQ ID NOs: 3-18, 23-26 (Table 2).

TABLE 2

Identified Aquaporin Genes

SEQ ID NO:				SEQ ID NO:
(Polynucleotide)	Gene name	Cluster name	Organism	(Polypeptide)

1	MAB54	tomato\|gb164\|BG125449	tomato	27
2	MAB55	tomato\|gb164\|BG134896	tomato	28
3	MAB56	tomato\|gb164\|AW218990	tomato	29
4	MAB57	tomato\|gb164\|AA824812	tomato		30
5	MAB58	tomato\|gb164\|BP881534	tomato	31
7	MAB69	tomato\|gb164\|AI637360	tomato	33
8	MAB70	tomato\|gb164\|BG133531	tomato	34
9	MAB71	tomato\|gb164\|BG629975	tomato	35
10	MAB72	tomato\|gb164\|BG136017	tomato	36
11	MAB73	tomato\|gb164\|BG131871	tomato	37
12	MAB74	tomato\|gb164\|AI775489	tomato	38
13	MAB75	tomato\|gb164\|BG136239	tomato	39
14	MAB76	tomato\|gb164\|BG134058	tomato	40
15	MAB77	tomato\|gb164\|BG629900	tomato		41
16	MAB78	tomato\|gb164\|BG130774	tomato	42
17	MAB79	tomato\|gb164\|BG124486	tomato	43
18	MAB80	tomato\|gb164\|AI483521	tomato	44
19	MAB81	tomato\|gb164\|CO751453	tomato	45
20	MAB115	barley\|gb157.2\|BF626376	barley	46
22	MAB117	barley\|gb157.2\|BE412516	barley	48
23	MAB119	tomato\|gb164\|BG134199	tomato	49
24	MAB176	tomato\|gb164\|CO635830	tomato	50
25	MAB177	tomato\|gb164\|CO751496	tomato	51
26	MAB178	tomato\|gb164\|CO751374	tomato	52

Table 2.

Sequences which are homologous [showing at least 80% protein sequence identity on 80% of the global hit or query length, as calculated using BlastP and tBlastN algorithms of the National Center of Biotechnology Information (NCBI)] or orthologues of the AQP genes described in Table 2, and are expected to possess the same role in ABST and yield improvement in plants, are disclosed in Table 3 hereinbelow (SEQ ID NOs:6, 215-1101 and 1138-1400; Table 3). In addition, Table 3 also includes homologous and orthologues of the AQP TIP2 subfamily (SEQ ID NOs:21, 53-214, 1102-1137) and additional homologous and orthologues (SEQ ID NOs:2844-3051).

TABLE 3

Polynucleotide and polypeptide sequences of AQP homologous and orthologous

Polynuc.			Polypep.	Hom.			%
SEQ ID			SEQ ID	of SEQ ID	%	Query	Subject
NO:	Cluster name	Organism	NO:	NO:	Ident.	cover.	cover.	Algorithm

53	apple\|gb157.3\|CN883304_T1	apple	1401	27	84	92.3387097	100	blastp
54	apple\|gb157.3\|CN489003_T1	apple	1402	27	83	100	100	blastp
55	aquilegia\|gb157.3\|	aquilegia	1403	27	85	100	100	blastp
	DR915168_T1
56	arabidopsis\|gb165\|	arabidopsis	1404	27	81	99.1935484	98.8	blastp
	AT3G16240_T1
57	artemisia\|gb164\|	artemisia	1405	27	80	98.7903226	99.1902834	blastp
	EY035829_T1
58	artemisia\|gb164\|	artemisia	1406	27	85	62.9032258	100	blastp
	EY113320_T1
59	artemisia\|gb164\|	artemisia	1407	27	81	98.7903226	99.1902834	blastp
	EY070770_T1
60	avocado\|gb164\|CV002132_T1	avocado	1408	27	84	50	100	blastp
61	b_juncea\|gb164\|	b_juncea	1409	27	83	100	100	blastp
	EVGN00333108491419_T1
62	b_juncea\|gb164\|	b_juncea	1410	27	82	53.6290323	97.0588235	blastp
	EVGN00503709641655_T1
63	b_juncea\|gb164\|	b_juncea	1411	27	84	63.7096774	100	blastp
	EVGN01003711220829_T1
64	b_oleracea\|gb161\|	b_oleracea	1412	27	84	100	100	blastp
	AM059585_T1
65	b_oleracea\|gb161\|	b_oleracea	1413	27	82	100	100	blastp
	AM385334_T1
66	b_oleracea\|gb161\|	b_oleracea	1414	27	81	100	100	blastp
	AM385915_T1
67	b_rapa\|gb162\|BG543171_T1	b_rapa	1415	27	82	100	100	blastp
68	b_rapa\|gb162\|BG543223_T1	b_rapa	1416	27	83	100	100	blastp
69	b_rapa\|gb162\|L37478_T1	b_rapa	1417	27	81	100	100	blastp
70	banana\|gb160\|DN238689_T1	banana	1418	27	83	76.6129032	100	blastp
71	bean\|gb164\|CB540614_T1	bean	1419	27	83	98.7903226	98.7903226	blastp
72	canola\|gb161\|EV092237_T1	canola	1420	27	84	100	100	blastp
73	canola\|gb161\|CN828178_T1	canola	1421	27	81	100	100	blastp
74	canola\|gb161\|CX188169_T1	canola	1422	27	82	100	100	blastp
75	canola\|gb161\|CD840590_T1	canola	1423	27	81	100	100	blastp
76	cassava\|gb164\|CK650415_T1	cassava	1424	27	85	100	100	blastp
77	castorbean\|gb160\|	castorbean	1425	27	87	100	100	blastp
	EE254645_T1
78	centaurea\|gb161\|	centaurea	1426	27	84	95.9677419	84.3971631	blastp
	EH725826_T1
79	centaurea\|gb161\|	centaurea	1427	27	88	89.1129032	97.7876106	blastp
	EL932474_T1
80	cherry\|gb157.2\|EE488049_T1	cherry	1428	27	80	69.3548387	100	blastp
81	cichorium\|gb161\|	cichorium	1429	27	80	100	92.8571429	blastp
	DT211633_T1
82	cichorium\|gb161\|	cichorium	1430	27	87	100	100	blastp
	EH672622_T1
83	citrus\|gb157.2\|CX663669_T1	citrus	1431	27	88	98.7903226	99.1902834	blastp
84	citrus\|gb157.2\|CF417983_T1	citrus	1432	27	88	98.7903226	99.1902834	blastp
85	citrus\|gb157.2\|CK665344_T1	citrus	1433	27	88	98.7903226	99.1902834	blastp
86	citrus\|gb157.2\|CK665344_T2	citrus	1434	27	87	82.2580645	100	blastp
87	clover\|gb162\|BB908328_T1	clover	1435	27	81	69.7580645	100	blastp
88	cotton\|gb164\|AF009567_T1	cotton	1436	27	87	100	100	blastp
89	cowpea\|gb166\|FF397761_T1	cowpea	1437	27	84	100	100	blastp
90	cowpea\|gb166\|FC457059_T1	cowpea	1438	27	83	98.7903226	98.7903226	blastp
91	dandelion\|gb161\|	dandelion	1439	27	85	100	100	blastp
	DY818755_T1
92	ginger\|gb164\|DY358186_T1	ginger	1440	27	80	98.3870968	99.1836735	blastp
93	ginger\|gb164\|DY351866_T1	ginger	1441	27	80	98.3870968	99.1836735	blastp
94	iceplant\|gb164\|AF133532_T1	iceplant	1442	27	81	100	100	blastp
95	ipomoea\|gb157.2\|	ipomoea	1443	27	87	100	100	blastp
	BJ576630_T1
96	lettuce\|gb157.2\|	lettuce	1444	27	87	100	100	blastp
	DW074363_T1
97	lettuce\|gb157.2\|	lettuce	1445	27	85	50.8064516	90.647482	blastp
	DW074363_T2
98	lettuce\|gb157.2\|	lettuce	1446	27	87	100	100	blastp
	DW145132_T1
99	lettuce\|gb157.2\|	lettuce	1447	27	87	100	100	blastp
	DW043760_T1
100	lettuce\|gb157.2\|	lettuce	1448	27	87	100	100	blastp
	DW104999_T1
101	lotus\|gb157.2\|BI420407_T1	lotus	1449	27	84	100	100	blastp
102	lotus\|gb157.2\|BF177457_T1	lotus	1450	27	86	98.7903226	98.7951807	blastp
103	medicago\|gb157.2\|	medicago	1451	27	83	89.1129032	94.8497854	blastp
	AI974300_T1
104	medicago\|gb157.2\|	medicago	1452	27	84	100	100	blastp
	AA660400_T1
105	nicotiana_benthamiana\|	nicotiana_benthamiana	1453	7	86	98.7903226	98.7903226	blastp
	gb162\|CN741988_T1
106	nicotiana_benthamiana\|	nicotiana_benthamiana	1454	27	92	100	100	blastp
	gb162\|CN655366_T1
107	nicotiana_benthamiana\|	nicotiana_benthamiana	1455	27	89	98.7903226	98.7903226	blastp
	gb162\|CN741998_T1
108	nicotiana_benthamiana\|	nicotiana_benthamiana	1456	27	93	100	100	blastp
	gb162\|CN742343_T1
109	peach\|gb157.2\|BU045214_T1	peach	1457	27	82	100	100	blastp
110	pepper\|gb157.2\|	pepper	1458	27	84	61.6935484	100	blastp
	CO776446_T1
111	pepper\|gb157.2\|	pepper	1459	27	86	63.3064516	100	blastp
	CA518313_T1
112	periwinkle\|gb164\|	periwinkle	1460	27	88	99.1935484	99.1935484	blastp
	EG555051_T1
113	petunia\|gb157.2\|	petunia	1461	27	84	63.7096774	85.2517986	tblastn
	CV296219_T1
114	poplar\|gb157.2\|BI129443_T1	poplar	1462	27	86	100	100	blastp
115	poplar\|gb157.2\|AI166943_T2	poplar	1463	27	83	61.6935484	100	blastp
116	poplar\|gb157.2\|AI166943_T1	poplar	1464	27	85	100	100	blastp
117	poplar\|gb157.2\|BI127662_T1	poplar	1465	27	89	79.0322581	100	blastp
118	potato\|gb157.2\|BQ513382_T1	potato	1466	27	97	100	100	blastp
119	potato\|gb157.2\|BQ513382_T2	potato	1467	27	97	50.8064516	96.1832061	blastp
120	radish\|gb164\|EV538411_T1	radish	1468	27	82	92.3387097	100	blastp
121	radish\|gb164\|AB010416_T1	radish	1469	27	81	100	100	blastp
122	radish\|gb164\|EW725945_T1	radish	1470	27	82	100	100	blastp
123	radish\|gb164\|EV527946_T1	radish	1471	27	81	100	100	blastp
124	rose\|gb157.2\|BQ104096_T1	rose	1472	27	85	85.0806452	95.045045	blastp
125	safflower\|gb162\|	safflower	1473	27	87	100	100	blastp
	EL374001_T1
126	safflower\|gb162\|	safflower	1474	27	83	100	100	blastp
	EL406178_T1
127	sesame\|gb157.2\|	sesame	1475	27	81	61.2903226	86.3636364	tblastn
	BU668161_T1
128	soybean\|gb166\|AW349399_T1	soybean	1476	27	84	98.7903226	98.7903226	blastp
129	soybean\|gb166\|CD416937_T1	soybean	1477	27	85	75.4032258	100	blastp
130	soybean\|gb166\|CA786095_T1	soybean	1478	27	84	98.7903226	98.7903226	blastp
131	soybean\|gb166\|AW350817_T1	soybean	1479	27	81	100	100	blastp
132	spruce\|gb162\|CO216479_T1	spruce	1480	27	80	98.7903226	98.4	blastp
133	strawberry\|gb164\|	strawberry	1481	27	82	100	100	blastp
	DV438565_T1
134	sunflower\|gb162\|	sunflower	1482	27	82	100	100	blastp
	X95952_T1
135	sunflower\|gb162\|	sunflower	1483	27	83	100	100	blastp
	CD847513_T1
136	sunflower\|gb162\|	sunflower	1484	27	84	100	100	blastp
	CD845750_T1
137	sunflower\|gb162\|	sunflower	1485	27	86	100	100	blastp
	CD848081_T1
138	sunflower\|gb162\|	sunflower	1486	27	83	100	100	blastp
	CD849577_T1
139	tobacco\|gb162\|CV016921_T1	tobacco	1487	27	88	100	100	blastp
140	tobacco\|gb162\|CV018684_T1	tobacco	1488	27	93	100	100	blastp
141	tobacco\|gb162\|CV019641_T1	tobacco	1489	27	91	100	100	blastp
142	tomato\|gb164\|AW626247_T1	tomato	No	27	83	50	88.3610451	tblastn
			predicted
			protein
143	triphysaria\|gb164\|	triphysaria	1490	27	81	100	100	blastp
	EX999390_T1
144	triphysaria\|gb164\|	triphysaria	1491	27	80	100	100	blastp
	BM356478_T1
145	triphysaria\|gb164\|	triphysaria	1492	27	81	81.0483871	98.0487805	blastp
	BM356478_T2
146	aquilegia\|gb157.3\|	aquilegia	1493	28	86	100	100	blastp
	DR922172_T1
147	arabidopsis\|gb165\|	arabidopsis	1494	28	82	98.8	98.8	blastp
	AT5G47450_T1
148	arabidopsis\|gb165\|	arabidopsis	1495	28	85	99.6	99.6	blastp
	AT4G17340_T1
149	artemisia\|gb164\|	artemisia	1496	28	83	69.2	100	blastp
	EY080612_T1
150	b_rapa\|gb162\|EX104899_T1	b_rapa	1497	28	84	95.2	99.1666667	blastp
151	canola\|gb161\|EE430505_T1	canola	1498	28	85	95.2	94.8207171	blastp
152	canola\|gb161\|DY017904_T1	canola	1499	28	82	98.8	98.8	blastp
153	canola\|gb161\|EL590702_T1	canola	1500	28	84	99.6	99.6	blastp
154	canola\|gb161\|CD818320_T1	canola	1501	28	85	99.6	99.6	blastp
155	cassava\|gb164\|DB923860_T1	cassava	1502	28	81	100	100	blastp
156	centaurea\|gb161\|	centaurea	1503	28	84	98.8	99.1935484	blastp
	EL932179_T1
157	centaurea\|gb161\|	centaurea	1504	28	82	87.6	88.9795918	blastp
	EH718862_T1
158	cichorium\|gb161\|	cichorium	1505	28	86	88.8	64.7230321	tblastn
	EH689841_T1
159	citrus\|gb157.2\|CO913277_T1	citrus	1506	28	84	100	100	blastp
160	citrus\|gb157.2\|CO912449_T1	citrus	1507	28	82	95.6	75.2360965	tblastn
161	cotton\|gb164\|DV437956_T1	cotton	1508	28	88	50.8	100	blastp
162	cotton\|gb164\|CD486503_T1	cotton	1509	28	86	100	100	blastp
163	dandelion\|gb161\|	dandelion	1510	28	84	100	100	blastp
	DY827614_T1
164	dandelion\|gb161\|	dandelion	1511	28	86	100	100	blastp
	DY822865_T1
165	dandelion\|gb161\|	dandelion	1512	28	86	100	100	blastp
	DY819043_T1
166	iceplant\|gb164\|AF133533_T1	iceplant	1513	28	82	82	99.5145631	blastp
167	lettuce\|gb157.2\|	lettuce	1514	28	85	100	100	blastp
	DW057721_T1
168	lettuce\|gb157.2\|	lettuce	1515	28	85	100	100	blastp
	DW045203_T1
169	lettuce\|gb157.2\|	lettuce	1516	28	84	100	100	blastp
	DW046133_T1
170	lettuce\|gb157.2\|	lettuce	1517	28	85	85.6	100	blastp
	DW080742_T1
171	lettuce\|gb157.2\|	lettuce	1518	28	86	100	100	blastp
	DW075611_T1
172	lettuce\|gb157.2\|	lettuce	1519	28	86	100	100	blastp
	DW123899_T1
173	lettuce\|gb157.2\|	lettuce	1520	28	84	100	100	blastp
	DW103468_T1
174	lettuce\|gb157.2\|	lettuce	1521	28	85	100	100	blastp
	DW161237_T1
175	lettuce\|gb157.2\|	lettuce	1522	28	85	100	100	blastp
	DW079798_T1
176	lettuce\|gb157.2\|	lettuce	1523	28	85	100	100	blastp
	DW079554_T1
177	lettuce\|gb157.2\|	lettuce	1524	28	85	100	100	blastp
	DW147378_T1
178	lettuce\|gb157.2\|	lettuce	1525	28	83	100	100	blastp
	DW052373_T1
179	lettuce\|gb157.2\|	lettuce	1526	28	86	100	100	blastp
	DW105592_T1
180	lettuce\|gb157.2\|	lettuce	1527	28	85	100	100	blastp
	DW075384_T1
181	lettuce\|gb157.2\|	lettuce	1528	28	86	100	100	blastp
	DW155153_T1
182	lettuce\|gb157.2\|	lettuce	1529	28	85	100	100	blastp
	CV699993_T1
183	lettuce\|gb157.2\|	lettuce	1530	28	84	100	100	blastp
	DW078166_T1
184	lotus\|gb157.2\|AV409092_T1	lotus	1531	28	80	53.2	100	blastp
185	medicago\|gb157.2\|	medicago	1532	28	81	98.8	99.5951417	blastp
	AI974377_T1
186	melon\|gb165\|AM725511_T1	melon	1533	28	84	100	100	blastp
187	nicotiana_benthamiana\|	nicotiana_benthamiana	1534	28	90	70.4	100	blastp
	gb162\|EH370474_T1
188	onion\|gb162\|BE205571_T1	onion	1535	28	83	99.6	99.5967742	blastp
189	onion\|gb162\|AA601764_T1	onion	1536	28	86	95.2	96.3414634	blastp
190	papaya\|gb165\|EX255759_T1	papaya	1537	28	82	99.6	99.5983936	blastp
191	peanut\|gb161\|EH043676_T1	peanut	1538	28	82	99.6	99.5967742	blastp
192	pepper\|gb157.2\|	pepper	1539	28	94	86.4	100	blastp
	CK901741_T1
193	periwinkle\|gb164\|	periwinkle	1540	28	86	64.4	96.4071856	blastp
	FD423620_T1
194	poplar\|gb157.2\|BU886993_T1	poplar	1541	28	84	100	100	blastp
195	poplar\|gb157.2\|CA826065_T1	poplar	1542	28	85	86.8	100	blastp
196	potato\|gb157.2\|BG591546_T2	potato	1543	28	81	100	100	blastp
197	radish\|gb164\|EV531940_T1	radish	1544	28	83	94.8	94.8	blastp
198	radish\|gb164\|EV527785_T1	radish	1545	28	84	92.8	95.473251	blastp
199	radish\|gb164\|EV550763_T1	radish	1546	28	84	95.2	94.8207171	blastp
200	radish\|gb164\|EX902593_T1	radish	1547	28	85	95.2	99.58159	blastp
201	radish\|gb164\|EV535199_T1	radish	1548	28	84	96	98.7654321	blastp
202	radish\|gb164\|EV525705_T1	radish	1549	28	85	96	98.7654321	blastp
203	safflower\|gb162\|	safflower	1550	28	86	86.8	100	blastp
	EL399548_T1
204	safflower\|gb162\|	safflower	1551	28	87	100	100	blastp
	EL376421_T1
205	spurge\|gb161\|DV127241_T1	spurge	1552	28	85	88.4	99.103139	blastp
206	strawberry\|gb164\|	strawberry	1553	28	82	100	100	blastp
	GFXDQ178022X1_T1
207	sunflower\|gb162\|	sunflower	1554	28	86	100	100	blastp
	X95953_T1
208	sunflower\|gb162\|	sunflower	1555	28	85	100	100	blastp
	DY911049_T1
209	sunflower\|gb162\|	sunflower	1556	28	81	88.8	98.2300885	blastp
	DY921796_T1
210	sunflower\|gb162\|	sunflower	1557	28	85	100	100	blastp
	DY918762_T1
211	sunflower\|gb162\|	sunflower	1558	28	81	100	100	blastp
	DY906198_T1
212	sunflower\|gb162\|	sunflower	1559	28	81	100	100	blastp
	DY932268_T1
213	tobacco\|gb162\|GFXS45406X1_T1	tobacco	1560	28	93	100	100	blastp
214	tobacco\|gb162\|EB445911_T1	tobacco	1561	28	89	100	100	blastp
215	apricot\|gb157.2\|	apricot	1562	29	81	54.5454545	95.8333333	blastp
	CB818493_T1
216	arabidopsis\|gb165\|	arabidopsis	1563	29	80	99.2094862	99.6031746	blastp
	AT4G01470_T1
217	avocado\|gb164\|CK760396_T1	avocado	1564	29	81	54.5454545	93.877551	blastp
218	b_rapa\|gb162\|EX017183_T1	b_rapa	1565	29	83	69.5652174	94.1176471	blastp
219	barley\|gb157.3\|BE413237_T1	barley	1566	29	81	99.2094862	99.6031746	blastp
220	cassava\|gb164\|CK644827_T1	cassava	1567	29	90	57.312253	99.3150685	blastp
221	cassava\|gb164\|BM259770_T1	cassava	1568	29	82	99.2094862	99.6031746	blastp
222	cassava\|gb164\|CK645124_T1	cassava	1569	29	80	99.2094862	99.6031746	blastp
223	castorbean\|gb160\|	castorbean	1570	29	84	99.2094862	99.6031746	blastp
	EG666198_T1
224	castorbean\|gb160\|	castorbean	1571	29	82	99.2094862	99.6015936	blastp
	AJ605571_T1
225	castorbean\|gb160\|	castorbean	1572	29	83	99.2094862	99.6031746	blastp
	AJ605570_T1
226	centaurea\|gb161\|	centaurea	1573	29	88	74.7035573	97.9274611	blastp
	EL931525_T1
227	cichorium\|gb161\|	cichorium	1574	29	87	65.6126482	99.4011976	blastp
	EH707617_T1
228	citrus\|gb157.2\|CF834233_T1	citrus	1575	29	80	94.4664032	94.4444444	blastp
229	citrus\|gb157.2\|BQ624227_T1	citrus	1576	29	87	99.2094862	99.6031746	blastp
230	citrus\|gb157.2\|BQ623056_T1	citrus	1577	29	87	97.2332016	98.4	blastp
231	citrus\|gb157.2\|BQ624617_T1	citrus	1578	29	87	99.2094862	99.6031746	blastp
232	cotton\|gb164\|CD486523_T1	cotton	1579	29	81	99.2094862	99.6031746	blastp
233	cotton\|gb164\|AI729919_T1	cotton	1580	29	82	99.2094862	99.6031746	blastp
234	cotton\|gb164\|BG442315_T1	cotton	1581	29	86	99.2094862	99.6031746	blastp
235	cotton\|gb164\|EX167179_T1	cotton	1582	29	84	56.916996	100	blastp
236	cotton\|gb164\|AI726375_T1	cotton	1583	29	82	99.2094862	99.6031746	blastp
237	cowpea\|gb166\|FF384697_T1	cowpea	1584	29	84	99.2094862	99.6031746	blastp
238	dandelion\|gb161\|	dandelion	1585	29	88	99.2094862	99.6031746	blastp
	DY825779_T1
239	fescue\|gb161\|CK802772_T1	fescue	1586	29	80	99.2094862	99.6031746	blastp
240	grape\|gb160\|BQ796848_T1	grape	1587	29	84	99.2094862	99.6015936	blastp
241	grape\|gb160\|CF605030_T1	grape	1588	29	82	99.2094862	99.6031746	blastp
242	ipomoea\|gb157.2\|	ipomoea	1589	29	85	53.7549407	100	blastp
	EE883704_T1
243	ipomoea\|gb157.2\|	ipomoea	1590	29	85	99.2094862	99.6031746	blastp
	BJ554617_T1
244	lettuce\|gb157.2\|	lettuce	1591	29	87	99.2094862	99.6031746	blastp
	DW074608_T1
245	lettuce\|gb157.2\|	lettuce	1592	29	87	99.2094862	99.6031746	blastp
	DY977540_T1
246	lotus\|gb157.2\|BW615882_T1	lotus	1593	29	80	50.1976285	100	blastp
247	maize\|gb164\|DQ245749_T1	maize	1594	29	81	99.2094862	99.6031746	blastp
248	medicago\|gb157.2\|	medicago	1595	29	82	99.2094862	99.6031746	blastp
	BI266516_T1
249	nicotiana_benthamiana\|	nicotiana_benthamiana	1596	29	94	83.0039526	99.5260664	blastp
	gb162\|CN743053_T1
250	papaya\|gb165\|EX256526_T1	papaya	1597	29	80	99.2094862	99.6031746	blastp
251	papaya\|gb165\|EX255270_T1	papaya	1598	29	86	99.2094862	99.6031746	blastp
252	peach\|gb157.2\|AF367456_T1	peach	1599	29	84	53.7549407	100	blastp
253	pepper\|gb157.2\|	pepper	1600	29	81	73.1225296	98.9304813	blastp
	CK902019_T1
254	poplar\|gb157.2\|AI163470_T1	poplar	1601	29	81	99.2094862	99.6031746	blastp
255	poplar\|gb157.2\|AI166549_T1	poplar	1602	29	82	99.2094862	99.6031746	blastp
256	poplar\|gb157.2\|BU887722_T1	poplar	1603	29	81	99.2094862	99.6031746	blastp
257	poplar\|gb157.2\|BU875073_T1	poplar	1604	29	83	99.2094862	99.6031746	blastp
258	poplar\|gb157.2\|CA823737_T1	poplar	1605	29	88	53.3596838	99.2647059	blastp
259	poplar\|gb157.2\|AI166136_T1	poplar	1606	29	80	99.2094862	99.6031746	blastp
260	radish\|gb164\|EV544876_T1	radish	1607	29	80	99.2094862	99.6031746	blastp
261	rice\|gb157.2\|AA752956_T1	rice	1608	29	80	99.2094862	99.6031746	blastp
262	soybean\|gb166\|CX703984_T1	soybean	1609	29	80	99.2094862	99.6031746	blastp
263	soybean\|gb166\|SOYNODB_T1	soybean	1610	29	86	99.2094862	99.6031746	blastp
264	spurge\|gb161\|DV146067_T1	spurge	1611	29	84	87.3517787	100	blastp
265	spurge\|gb161\|AW990927_T1	spurge	1612	29	80	99.2094862	99.6031746	blastp
266	sunflower\|gb162\|	sunflower	1613	29	86	99.2094862	99.6031746	blastp
	DY919534_T1
267	tobacco\|gb162\|EB443312_T1	tobacco	1614	29	92	99.2094862	99.6031746	blastp
268	tobacco\|gb162\|CV019217_T1	tobacco	1615	29	81	98.0237154	99.5967742	blastp
269	tobacco\|gb162\|EB443618_T1	tobacco	1616	29	92	99.2094862	99.6031746	blastp
270	tobacco\|gb162\|CV018899_T1	tobacco	1617	29	81	98.0237154	99.5967742	blastp
271	wheat\|gb164\|BE418306_T1	wheat	1618	29	80	99.2094862	99.6031746	blastp
272	wheat\|gb164\|BE404792_T1	wheat	1619	29	82	52.9644269	99.2592593	blastp
273	wheat\|gb164\|BE216922_T1	wheat	1620	29	81	99.2094862	99.6031746	blastp
274	artemisia\|gb164\|	artemisia	1621	30	80	79.6	100	blastp
	EY083433_T1
275	banana\|gb160\|ES432704_T1	banana	1622	30	81	88.8	93.6708861	blastp
276	banana\|gb160\|DN238541_T1	banana	1623	30	80	100	100	blastp
277	barley\|gb157.3\|BE412510_T1	barley	1624	30	80	100	100	blastp
278	cotton\|gb164\|AI726168_T1	cotton	1625	30	80	98.8	99.5983936	blastp
279	cotton\|gb164\|AI731742_T1	cotton	1626	30	80	100	100	blastp
280	cotton\|gb164\|AI055329_T1	cotton	1627	30	80	100	100	blastp
281	grape\|gb160\|BQ794219_T1	grape	1628	30	81	100	100	blastp
282	ipomoea\|gb157.2\|	ipomoea	1629	30	81	98	100	blastp
	BJ554855_T1
283	ipomoea\|gb157.2\|	ipomoea	1630	30	84	100	100	blastp
	BM878761_T1
284	lettuce\|gb157.2\|	lettuce	1631	30	80	86.8	98.1981982	blastp
	DW045084_T1
285	lettuce\|gb157.2\|	lettuce	1632	30	80	88	99.103139	blastp
	DW114621_T1
286	lettuce\|gb157.2\|	lettuce	1633	30	81	50.8	100	blastp
	DW078778_T1
287	maize\|gb164\|CO528320_T1	maize	1634	30	82	69.6	100	blastp
288	maize\|gb164\|AW257922_T1	maize	1635	30	80	52.4	100	blastp
289	maize\|gb164\|BI675058_T1	maize	1636	30	80	54	99.2592593	blastp
290	maize\|gb164\|AW352518_T1	maize	1637	30	80	51.2	100	blastp
291	maize\|gb164\|AF037061_T1	maize	1638	30	81	100	100	blastp
292	nicotiana_benthamiana\|	nicotiana_benthamiana	1639	30	90	100	100	blastp
	gb162\|CN655062_T1
293	nicotiana_benthamiana\|	nicotiana_benthamiana	1640	30	90	100	100	blastp
	gb162\|CN741621_T1
294	oil_palm\|gb166\|	oil_palm	1641	30	80	100	100	blastp
	CN599861_T1
295	papaya\|gb165\|EX246150_T1	papaya	1642	30	82	100	100	blastp
296	pepper\|gb157.2\|	pepper	1643	30	91	99.2	98.8	blastp
	BM060520_T1
297	periwinkle\|gb164\|	periwinkle	1644	30	82	100	100	blastp
	EG554262_T1
298	petunia\|gb157.2\|	petunia	1645	30	89	100	100	blastp
	AF452015_T1
299	potato\|gb157.2\|CK853059_T1	potato	1646	30	96	92	91.3043478	blastp
300	potato\|gb157.2\|CK852742_T1	potato	1647	30	82	60.4	100	blastp
301	potato\|gb157.2\|BM407759_T1	potato	1648	30	99	81.2	97.5961538	blastp
302	potato\|gb157.2\|CK718033_T1	potato	1649	30	98	65.2	92.0903955	blastp
303	potato\|gb157.2\|CK717899_T1	potato	1650	30	100	62	95.0920245	blastp
304	potato\|gb157.2\|CV472240_T1	potato	1651	30	97	79.6	95.215311	blastp
305	potato\|gb157.2\|BG098199_T1	potato	1652	30	95	93.2	99.5726496	blastp
306	rice\|gb157.2\|U37925_T1	rice	1653	30	81	100	100	blastp
307	rye\|gb164\|BE494266_T1	rye	1654	30	80	100	100	blastp
308	sorghum\|gb161.xeno\|	sorghum	1655	30	80	100	100	blastp
	AF037061_T1
309	sugarcane\|gb157.2\|	sugarcane	1656	30	80	80.4	100	blastp
	BQ535365_T1
310	switchgrass\|gb165\|	switchgrass	1657	30	81	100	100	blastp
	DN141449_T1
311	switchgrass\|gb165\|	switchgrass	1658	30	81	100	100	blastp
	DN142089_T1
312	tobacco\|gb162\|CN824866_T1	tobacco	1659	30	90	100	100	blastp
313	tobacco\|gb162\|CV017118_T1	tobacco	1660	30	90	100	100	blastp
314	wheat\|gb164\|TAU86762_T1	wheat	1661	30	80	100	100	blastp
315	wheat\|gb164\|BE499589_T1	wheat	1662	30	80	72	100	blastp
316	lettuce\|gb157.2\|	lettuce	1663	31	80	100	100	blastp
	DW087170_T1
317	tobacco\|gb162\|EH616288_T1	tobacco	1664	32	87	50.7692308	85.1612903	blastp
318	apple\|gb157.3\|CO068608_T1	apple	1665	33	86	98.2638889	98.951049	blastp
319	apple\|gb157.3\|AB100869_T1	apple	1666	33	83	98.2638889	99.3079585	blastp
320	apple\|gb157.3\|AB100870_T1	apple	1667	33	83	98.2638889	99.3079585	blastp
321	apple\|gb157.3\|CN860225_T1	apple	1668	33	86	54.8611111	90.2857143	blastp
322	apple\|gb157.3\|CK900645_T1	apple	1669	33	85	98.2638889	98.951049	blastp
323	apricot\|gb157.2\|	apricot	1670	33	89	51.0416667	98.6577181	blastp
	CB822297_T1
324	apricot\|gb157.2\|	apricot	1671	33	83	98.2638889	98.9655172	blastp
	CB819647_T1
325	aquilegia\|gb157.3\|	aquilegia	1672	33	86	98.2638889	98.9547038	blastp
	DR917005_T1
326	arabidopsis\|gb165\|	arabidopsis	1673	33	84	73.6111111	97.260274	blastp
	AT4G00430_T2
327	arabidopsis\|gb165\|	arabidopsis	1674	33	86	88.1944444	84.3853821	blastp
	AT2G45960_T3
328	arabidopsis\|gb165\|	arabidopsis	1675	33	87	98.2638889	98.9547038	blastp
	AT4G23400_T1
329	arabidopsis\|gb165\|	arabidopsis	1676	33	86	98.2638889	98.951049	blastp
	AT2G45960_T1
330	arabidopsis\|gb165\|	arabidopsis	1677	33	87	98.2638889	98.9547038	blastp
	AT4G00430_T1
331	arabidopsis\|gb165\|	arabidopsis	1678	33	88	98.2638889	98.951049	blastp
	AT1G01620_T1
332	arabidopsis\|gb165\|	arabidopsis	1679	33	85	98.2638889	98.951049	blastp
	AT3G61430_T1
333	arabidopsis\|gb165\|	arabidopsis	1680	33	86	88.1944444	92.7007299	blastp
	AT2G45960_T4
334	artemisia\|gb164\|	artemisia	1681	33	86	98.2638889	98.9547038	blastp
	EY046087_T1
335	artemisia\|gb164\|	artemisia	1682	33	87	73.6111111	99.0697674	blastp
	EY046310_T1
336	artemisia\|gb164\|	artemisia	1683	33	84	97.5694444	98.2758621	blastp
	EY032836_T1
337	artemisia\|gb164\|	artemisia	1684	33	84	89.2361111	99.2307692	blastp
	EY031810_T1
338	avocado\|gb164\|CK751385_T1	avocado	1685	33	86	98.2638889	98.9547038	blastp
339	avocado\|gb164\|CK745633_T1	avocado	1686	33	91	73.6111111	99.0654206	blastp
340	b_juncea\|gb164\|	b_juncea	1687	33	87	98.2638889	98.951049	blastp
	EVGN00081008450640_T1
341	b_juncea\|gb164\|	b_juncea	1688	33	90	60.4166667	96.1325967	blastp
	EVGN00515811862066_T1
342	b_juncea\|gb164\|	b_juncea	1689	33	89	73.6111111	99.0654206	blastp
	EVGN00230716760965_T1
343	b_juncea\|gb164\|	b_juncea	1690	33	84	66.3194444	96.4646465	blastp
	EVGN01776308261252_T1
344	b_juncea\|gb164\|	b_juncea	1691	33	91	65.2777778	98.9473684	blastp
	EVGN00910030360678_T1
345	b_juncea\|gb164\|	b_juncea	1692	33	88	51.0416667	98.6577181	blastp
	EVGN03812526911787_T1
346	b_juncea\|gb164\|	b_juncea	1693	33	91	65.2777778	98.9473684	blastp
	EVGN00227203510305_T1
347	b_juncea\|gb164\|	b_juncea	1694	33	87	98.2638889	98.951049	blastp
	EVGN00462518410866_T1
348	b_juncea\|gb164\|	b_juncea	1695	33	89	73.2638889	99.0610329	blastp
	EVGN00248411120906_T1
349	b_juncea\|gb164\|	b_juncea	1696	33	86	98.2638889	98.2638889	blastp
	EF471211_T1
350	b_juncea\|gb164\|	b_juncea	1697	33	86	98.2638889	98.951049	blastp
	EVGN00440012650683_T1
351	b_juncea\|gb164\|	b_juncea	1698	33	86	98.2638889	98.951049	blastp
	EVGN00452211183349_T1
352	b_juncea\|gb164\|	b_juncea	1699	33	89	57.6388889	93.258427	blastp
	EVGN03595331210044_T1
353	b_juncea\|gb164\|	b_juncea	1700	33	87	98.2638889	98.951049	blastp
	EVGN00088009631302_T1
354	b_juncea\|gb164\|	b_juncea	1701	33	85	51.7361111	93.907563	tblastn
	EVGN00512912541009_T1
355	b_juncea\|gb164\|	b_juncea	1702	33	84	69.0972222	90.3177005	tblastn
	EVGN00756014550623_T1
356	b_oleracea\|gb161\|	b_oleracea	1703	33	86	98.2638889	98.951049	blastp
	AF299051_T1
357	b_oleracea\|gb161\|	b_oleracea	1704	33	87	75.3472222	99.5412844	blastp
	AM391520_T1
358	b_oleracea\|gb161\|	b_oleracea	1705	33	87	98.2638889	98.951049	blastp
	AM058918_T1
359	b_oleracea\|gb161\|	b_oleracea	1706	33	86	98.2638889	98.951049	blastp
	AF299050_T1
360	b_oleracea\|gb161\|	b_oleracea	1707	33	86	98.2638889	98.951049	blastp
	DY029936_T1
361	b_oleracea\|gb161\|	b_oleracea	1708	33	87	78.4722222	100	blastp
	EH422530_T1
362	b_rapa\|gb162\|CV546930_T1	b_rapa	1709	33	84	71.5277778	99.5169082	blastp
363	b_rapa\|gb162\|BG544387_T1	b_rapa	1710	33	86	95.4861111	99.2779783	blastp
364	b_rapa\|gb162\|CA992432_T1	b_rapa	1711	33	86	98.2638889	98.951049	blastp
365	b_rapa\|gb162\|CV546129_T2	b_rapa	1712	33	83	54.8611111	99.3710692	blastp
366	b_rapa\|gb162\|EE526280_T1	b_rapa	1713	33	87	98.2638889	98.951049	blastp
367	b_rapa\|gb162\|L33552_T1	b_rapa	1714	33	86	98.2638889	98.951049	blastp
368	b_rapa\|gb162\|BG543719_T1	b_rapa	1715	33	84	77.0833333	99.5515695	blastp
369	b_rapa\|gb162\|AF004293_T1	b_rapa	1716	33	87	98.2638889	98.951049	blastp
370	b_rapa\|gb162\|BG544086_T1	b_rapa	1717	33	87	98.2638889	98.951049	blastp
371	b_rapa\|gb162\|CX267412_T1	b_rapa	1718	33	86	98.2638889	99.2982456	blastp
372	b_rapa\|gb162\|CV546129_T1	b_rapa	1719	33	86	98.2638889	98.2638889	blastp
373	b_rapa\|gb162\|CV545634_T1	b_rapa	1720	33	83	56.5972222	90.5555556	blastp
374	banana\|gb160\|DN238827_T1	banana	1721	33	84	60.7638889	99.4285714	blastp
375	banana\|gb160\|ES431094_T1	banana	1722	33	89	63.8888889	98.9247312	blastp
376	barley\|gb157.3\|BE412959_T2	barley	1723	33	83	98.2638889	98.9726027	blastp
377	barley\|gb157.3\|AL507831_T1	barley	1724	33	85	99.3055556	99.6551724	blastp
378	barley\|gb157.3\|BE412959_T1	barley	1725	33	83	98.2638889	98.9726027	blastp
379	barley\|gb157.3\|BE412959_T5	barley	1726	33	82	98.2638889	98.9830508	blastp
380	barley\|gb157.3\|BE412959_T4	barley	1727	33	82	98.2638889	98.9830508	blastp
381	barley\|gb157.3\|AL502020_T1	barley	1728	33	82	98.2638889	98.9726027	blastp
382	barley\|gb157.3\|BE412972_T1	barley	1729	33	85	98.2638889	98.9583333	blastp
383	basilicum\|gb157.3\|	basilicum	1730	33	88	61.8055556	99.4413408	blastp
	DY340092_T1
384	basilicum\|gb157.3\|	basilicum	1731	33	87	73.9583333	99.5327103	blastp
	DY332264_T1
385	bean\|gb164\|CB543592_T1	bean	1732	33	88	98.2638889	98.9547038	blastp
386	bean\|gb164\|PVU97023_T1	bean	1733	33	84	98.2638889	99.3079585	blastp
387	bean\|gb164\|CB542193_T1	bean	1734	33	84	98.6111111	99.6539792	blastp
388	beet\|gb162\|BVU60149_T1	beet	1735	33	84	98.2638889	98.951049	blastp
389	brachypodium\|gb161.xeno\|	brachypodium	1736	33	83	82.9861111	95.6521739	blastp
	BE443278_T1
390	brachypodium\|gb161.xeno\|	brachypodium	1737	33	85	98.2638889	98.9583333	blastp
	BE216990_T1
391	brachypodium\|gb161.xeno\|	brachypodium	1738	33	82	86.8055556	72.7011494	blastp
	BE403307_T1
392	canola\|gb161\|CN731957_T1	canola	1739	33	86	98.2638889	98.951049	blastp
393	canola\|gb161\|CX194503_T1	canola	1740	33	86	98.2638889	98.951049	blastp
394	canola\|gb161\|CD814405_T1	canola	1741	33	87	98.2638889	98.951049	blastp
395	canola\|gb161\|EG020906_T1	canola	1742	33	86	98.2638889	98.951049	blastp
396	canola\|gb161\|H74720_T1	canola	1743	33	86	98.2638889	98.951049	blastp
397	canola\|gb161\|CN831315_T1	canola	1744	33	86	84.0277778	93.4362934	blastp
398	canola\|gb161\|CD817408_T1	canola	1745	33	87	98.2638889	98.951049	blastp
399	canola\|gb161\|EE502121_T1	canola	1746	33	89	52.7777778	98.7096774	blastp
400	canola\|gb161\|CX187544_T1	canola	1747	33	87	98.2638889	98.951049	blastp
401	canola\|gb161\|CD822064_T1	canola	1748	33	86	98.2638889	98.951049	blastp
402	canola\|gb161\|CD824965_T1	canola	1749	33	81	92.0138889	93.0313589	blastp
403	canola\|gb161\|EE485551_T1	canola	1750	33	87	98.2638889	98.951049	blastp
404	canola\|gb161\|CB686274_T1	canola	1751	33	86	98.2638889	98.951049	blastp
405	canola\|gb161\|CD814573_T1	canola	1752	33	83	76.3888889	94.0425532	blastp
406	canola\|gb161\|CX193398_T1	canola	1753	33	86	98.2638889	98.2638889	blastp
407	canola\|gb161\|CD818853_T1	canola	1754	33	86	98.2638889	98.951049	blastp
408	canola\|gb161\|DY005979_T1	canola	1755	33	85	78.4722222	99.5594714	blastp
409	canola\|gb161\|EE464964_T1	canola	1756	33	85	81.5972222	99.5762712	blastp
410	cassava\|gb164\|BM260264_T1	cassava	1757	33	85	97.9166667	99.3031359	blastp
411	cassava\|gb164\|CK901165_T1	cassava	1758	33	87	73.6111111	99.0697674	blastp
412	cassava\|gb164\|CK642415_T1	cassava	1759	33	87	98.2638889	98.9547038	blastp
413	cassava\|gb164\|DV455398_T1	cassava	1760	33	85	97.9166667	99.3031359	blastp
414	castorbean\|gb160\|	castorbean	1761	33	89	98.2638889	98.9547038	blastp
	T14819_T1
415	castorbean\|gb160\|	castorbean	1762	33	86	98.2638889	98.951049	blastp
	EG691229_T1
416	castorbean\|gb160\|	castorbean	1763	33	87	97.9166667	99.3031359	blastp
	AJ605566_T1
417	castorbean\|gb160\|	castorbean	1764	33	84	98.2638889	99.3055556	blastp
	MDL29969M000266_T1
418	castorbean\|gb160\|	castorbean	1765	33	87	97.9166667	98.9583333	blastp
	AJ605574_T1
419	centaurea\|gb161\|	centaurea	1766	33	82	97.5694444	98.6062718	blastp
	EH732068_T1
420	cichorium\|gb161\|	cichorium	1767	33	87	98.2638889	98.9547038	blastp
	EH673032_T1
421	cichorium\|gb161\|	cichorium	1768	33	90	51.7361111	98.6842105	blastp
	EH706808_T1
422	cichorium\|gb161\|	cichorium	1769	33	86	64.9305556	95.4314721	blastp
	EH701938_T1
423	citrus\|gb157.2\|CO912471_T1	citrus	1770	33	82	69.4444444	99.5098039	blastp
424	citrus\|gb157.2\|BQ624312_T1	citrus	1771	33	88	98.2638889	98.9547038	blastp
425	citrus\|gb157.2\|CN182376_T1	citrus	1772	33	84	92.7083333	94.0559441	blastp
426	citrus\|gb157.2\|CB291370_T1	citrus	1773	33	89	98.2638889	98.9547038	blastp
427	citrus\|gb157.2\|CF833327_T1	citrus	1774	33	85	98.2638889	99.3031359	blastp
428	citrus\|gb157.2\|BQ624860_T1	citrus	1775	33	85	97.9166667	99.6503497	blastp
429	citrus\|gb157.2\|CF508404_T1	citrus	1776	33	81	97.9166667	99.6503497	blastp
430	citrus\|gb157.2\|CB293694_T1	citrus	1777	33	84	98.2638889	99.3031359	blastp
431	citrus\|gb157.2\|BQ622975_T1	citrus	1778	33	85	97.9166667	99.6503497	blastp
432	citrus\|gb157.2\|BE213453_T1	citrus	1779	33	86	73.6111111	99.5305164	blastp
433	citrus\|gb157.2\|CF828110_T1	citrus	1780	33	85	98.6111111	99.6527778	blastp
434	citrus\|gb157.2\|BQ623397_T1	citrus	1781	33	86	93.4027778	99.6336996	blastp
435	clover\|gb162\|BB903117_T1	clover	1782	33	85	98.6111111	99.6539792	blastp
436	coffea\|gb157.2\|BQ449035_T1	coffea	1783	33	88	98.9583333	99.3055556	blastp
437	coffea\|gb157.2\|DV663743_T1	coffea	1784	33	85	98.2638889	98.9473684	blastp
438	cotton\|gb164\|CD486529_T1	cotton	1785	33	83	98.2638889	99.3055556	blastp
439	cotton\|gb164\|BE052445_T1	cotton	1786	33	87	98.2638889	98.9547038	blastp
440	cotton\|gb164\|AI726690_T1	cotton	1787	33	86	98.2638889	98.9547038	blastp
441	cotton\|gb164\|DN803576_T1	cotton	1788	33	83	87.8472222	98.828125	blastp
442	cotton\|gb164\|BM358242_T1	cotton	1789	33	81	98.2638889	99.2882562	blastp
443	cotton\|gb164\|CO085369_T1	cotton	1790	33	81	52.7777778	99.3506494	blastp
444	cotton\|gb164\|CO098674_T1	cotton	1791	33	84	98.2638889	99.3055556	blastp
445	cotton\|gb164\|CO070796_T1	cotton	1792	33	84	98.2638889	99.3031359	blastp
446	cotton\|gb164\|AI729945_T1	cotton	1793	33	85	98.2638889	99.3079585	blastp
447	cotton\|gb164\|DW496760_T1	cotton	1794	33	86	80.5555556	98.3122363	blastp
448	cowpea\|gb166\|FF384916_T1	cowpea	1795	33	82	98.2638889	99.3031359	blastp
449	cowpea\|gb166\|FF555791_T1	cowpea	1796	33	82	98.6111111	99.6539792	blastp
450	cowpea\|gb166\|FC457489_T1	cowpea	1797	33	87	98.2638889	98.9547038	blastp
451	cowpea\|gb166\|AB037241_T1	cowpea	1798	33	84	98.2638889	99.3079585	blastp
452	cryptomeria\|gb166\|	cryptomeria	1799	33	84	71.5277788	99.5215311	blastp
	DC429824_T1
453	cryptomeria\|gb166\|	cryptomeria	1800	33	85	98.6111111	99.6515679	blastp
	AU036730_T1
454	dandelion\|gb161\|	dandelion	1801	33	84	82.2916667	93.7007874	blastp
	DY814032_T1
455	dandelion\|gb161\|	dandelion	1802	33	82	98.2638889	99.3031359	blastp
	DY802714_T1
456	dandelion\|gb161\|	dandelion	1803	33	84	95.8333333	99.6415771	blastp
	DY822683_T1
457	dandelion\|gb161\|	dandelion	1804	33	87	98.2638889	98.9583333	blastp
	DY806788_T1
458	dandelion\|gb161\|	dandelion	1805	33	84	84.0277778	99.5918367	blastp
	DY808781_T1
459	dandelion\|gb161\|	dandelion	1806	33	81	76.3888889	94.8497854	blastp
	DY810613_T1
460	fescue\|gb161\|CK803261_T1	fescue	1807	33	85	97.9166667	98.6111111	blastp
461	fescue\|gb161\|DT682664_T1	fescue	1808	33	84	67.3611111	99.4923858	blastp
462	fescue\|gb161\|DT677062_T1	fescue	1809	33	83	98.2638889	98.9655172	blastp
463	fescue\|gb161\|DT679061_T1	fescue	1810	33	86	98.2638889	98.9619377	blastp
464	flax\|gb157.3\|CV478314_T1	flax	1811	33	84	71.875	99.5260664	blastp
465	ginger\|gb164\|DY345344_T1	ginger	1812	33	88	98.2638889	98.951049	blastp
466	ginger\|gb164\|DY358322_T1	ginger	1813	33	85	98.2638889	98.9473684	blastp
467	ginger\|gb164\|DY360757_T1	ginger	1814	33	84	98.6111111	99.6478873	blastp
468	ginger\|gb164\|DY345596_T1	ginger	1815	33	86	98.2638889	98.9473684	blastp
469	grape\|gb160\|AF188844_T1	grape	1816	33	87	98.2638889	98.9547038	blastp
470	grape\|gb160\|AF188843_T1	grape	1817	33	85	98.2638889	98.951049	blastp
471	grape\|gb160\|AF188843_T3	grape	1818	33	85	98.2638889	98.951049	blastp
472	grape\|gb160\|CB971128_T1	grape	1819	33	87	98.2638889	99.3006993	blastp
473	grape\|gb160\|AF188843_T4	grape	1820	33	84	72.2222222	86.3070539	blastp
474	iceplant\|gb164\|	iceplant	1821	33	85	98.2638889	98.9473684	blastp
	MCU26537_T1
475	iceplant\|gb164\|CIPMIPA_T1	iceplant	1822	33	85	98.2638889	99.2957746	blastp
476	iceplant\|gb164\|CIPMIPB_T1	iceplant	1823	33	87	98.2638889	98.9473684	blastp
477	ipomoea\|gb157.2\|	ipomoea	1824	33	87	98.9583333	99.3031359	blastp
	BM878883_T1
478	ipomoea\|gb157.2\|	ipomoea	1825	33	85	98.6111111	99.6491228	blastp
	BJ553988_T1
479	ipomoea\|gb157.2\|	ipomoea	1826	33	84	98.6111111	99.6478873	blastp
	BJ553369_T1
480	ipomoea\|gb157.2\|	ipomoea	1827	33	89	98.9583333	99.3031359	blastp
	BJ553198_T1
481	lettuce\|gb157.2\|	lettuce	1828	33	86	98.2638889	97.9310345	blastp
	DW079915_T1
482	lettuce\|gb157.2\|	lettuce	1829	33	87	98.2638889	98.9547038	blastp
	DW043941_T1
483	lettuce\|gb157.2\|	lettuce	1830	33	87	96.875	98.245614	blastp
	DW047538_T1
484	lettuce\|gb157.2\|	lettuce	1831	33	84	98.2638889	99.3031359	blastp
	DW104582_T1
485	lettuce\|gb157.2\|	lettuce	1832	33	84	98.2638889	99.3031359	blastp
	DW044606_T1
486	lettuce\|gb157.2\|	lettuce	1833	33	85	89.2361111	94.1605839	blastp
	DW148209_T1
487	lettuce\|gb157.2\|	lettuce	1834	33	83	98.6111111	95.3333333	blastp
	DW148478_T1
488	lettuce\|gb157.2\|	lettuce	1835	33	86	98.2638889	98.9547038	blastp
	DW108503_T1
489	lettuce\|gb157.2\|	lettuce	1836	33	86	96.875	99.6441281	blastp
	DW046100_T1
490	lettuce\|gb157.2\|	lettuce	1837	33	85	98.2638889	99.3031359	blastp
	DW075079_T1
491	lettuce\|gb157.2\|	lettuce	1838	33	87	98.2638889	98.9547038	blastp
	DW076402_T1
492	lettuce\|gb157.2\|	lettuce	1839	33	86	89.5833333	92.8315412	blastp
	DW084041_T1
493	lettuce\|gb157.2\|	lettuce	1840	33	87	98.2638889	98.9547038	blastp
	DW145601_T1
494	lettuce\|gb157.2\|	lettuce	1841	33	86	98.2638889	98.9547038	blastp
	CV699980_T1
495	lettuce\|gb157.2\|	lettuce	1842	33	87	98.2638889	98.9547038	blastp
	DW064849_T1
496	lettuce\|gb157.2\|	lettuce	1843	33	83	98.6111111	89.0965732	blastp
	DW147179_T1
497	lettuce\|gb157.2\|	lettuce	1844	33	83	98.6111111	97.2789116	blastp
	DW045991_T1
498	lotus\|gb157.2\|AF145707_T1	lotus	1845	33	84	98.2638889	99.3079585	blastp
499	lotus\|gb157.2\|AI967594_T1	lotus	1846	33	86	96.875	98.245614	blastp
500	lotus\|gb157.2\|AF145708_T1	lotus	1847	33	87	66.6666667	100	blastp
501	maize\|gb164\|EC881658_T1	maize	1848	33	86	54.5138889	98.7421384	blastp
502	maize\|gb164\|AI372377_T1	maize	1849	33	85	98.2638889	98.9583333	blastp
503	maize\|gb164\|AF145706_T1	maize	1850	33	83	60.7638889	100	blastp
504	maize\|gb164\|AI855222_T1	maize	1851	33	84	98.2638889	98.9726027	blastp
505	maize\|gb164\|AI619392_T1	maize	1852	33	86	98.2638889	98.9619377	blastp
506	maize\|gb164\|AI861086_T1	maize	1853	33	84	98.2638889	98.9583333	blastp
507	medicago\|gb157.2\|	medicago	1854	33	84	98.2638889	99.3079585	blastp
	AW684000_T1
508	medicago\|gb157.2\|	medicago	1855	33	83	98.2638889	99.3079585	blastp
	AI974398_T1
509	medicago\|gb157.2\|	medicago	1856	33	88	97.5694444	98.6062718	blastp
	AL366983_T1
510	medicago\|gb157.2\|	medicago	1857	33	84	98.6111111	99.3103448	blastp
	AI737528_T1
511	medicago\|gb157.2\|	medicago	1858	33	84	82.2916667	87.2262774	blastp
	BQ151876_T1
512	melon\|gb165\|DV632745_T1	melon	1859	33	84	98.2638889	99.3150685	blastp
513	melon\|gb165\|CF674915_T1	melon	1860	33	83	98.2638889	99.3150685	blastp
514	melon\|gb165\|DV632772_T1	melon	1861	33	85	98.2638889	98.951049	blastp
515	millet\|gb161\|CD724341_T1	millet	1862	33	83	57.2916667	92.1787709	blastp
516	nicotiana_benthamiana\|	nicotiana_benthamiana	1863	33	84	66.6666667	97.9487179	blastp
	gb162\|ES885295_T1
517	oil_palm\|gb166\|	oil_palm	1864	33	85	98.2638889	98.9547038	blastp
	CN600863_T1
518	oil_palm\|gb166\|	oil_palm	1865	33	86	98.2638889	98.9547038	blastp
	CN600797_T1
519	onion\|gb162\|AF255796_T1	onion	1866	33	86	98.2638889	98.9583333	blastp
520	papaya\|gb165\|EX228092_T1	papaya	1867	33	84	72.2222222	93.2735426	blastp
521	papaya\|gb165\|AJ000031_T1	papaya	1868	33	85	98.2638889	99.3079585	blastp
522	papaya\|gb165\|EX257869_T1	papaya	1869	33	83	98.2638889	99.3031359	blastp
523	papaya\|gb165\|AM903842_T1	papaya	1870	33	90	98.2638889	98.951049	blastp
524	peach\|gb157.2\|BU039203_T1	peach	1871	33	84	98.2638889	98.9655172	blastp
525	peach\|gb157.2\|BU040913_T1	peach	1872	33	90	51.3888889	98.6666667	blastp
526	peanut\|gb161\|CD038184_T1	peanut	1873	33	83	98.6111111	99.6539792	blastp
527	peanut\|gb161\|ES490696_T1	peanut	1874	33	82	73.9583333	99.5348837	blastp
528	peanut\|gb161\|CD038104_T1	peanut	1875	33	83	98.2638889	99.3079585	blastp
529	pepper\|gb157.2\|	pepper	1876	33	97	96.875	100	blastp
	CA523071_T1
530	pepper\|gb157.2\|	pepper	1877	33	95	99.3055556	100	blastp
	BM063708_T1
531	periwinkle\|gb164\|	periwinkle	1878	33	88	98.9583333	99.3031359	blastp
	EG554502_T1
532	periwinkle\|gb164\|	periwinkle	1879	33	87	98.9583333	99.3031359	blastp
	EG554518_T1
533	periwinkle\|gb164\|	periwinkle	1880	33	83	84.0277778	96.8	blastp
	EG556773_T1
534	petunia\|gb157.2\|	petunia	1881	33	93	99.3055556	100	blastp
	AF452010_T1
535	petunia\|gb157.2\|	petunia	1882	33	92	61.8055556	100	blastp
	CV292775_T1
536	petunia\|gb157.2\|	petunia	1883	33	86	98.2638889	99.3006993	blastp
	AF452011_T1
537	pine\|gb157.2\|AL751335_T1	pine	1884	33	83	98.2638889	98.9583333	blastp
538	pine\|gb157.2\|AA556193_T1	pine	1885	33	85	98.2638889	98.9583333	blastp
539	pine\|gb157.2\|AL750485_T1	pine	1886	33	85	98.2638889	98.9583333	blastp
540	pineapple\|gb157.2\|	pineapple	1887	33	83	98.2638889	97.9452055	blastp
	DT335964_T1
541	pineapple\|gb157.2\|	pineapple	1888	33	86	98.2638889	98.9583333	blastp
	DT338557_T1
542	poplar\|gb157.2\|BU817536_T1	poplar	1889	33	83	98.2638889	99.3031359	blastp
543	poplar\|gb157.2\|AI162483_T1	poplar	1890	33	88	98.2638889	98.9583333	blastp
544	poplar\|gb157.2\|BI122420_T1	poplar	1891	33	87	97.9166667	99.3031359	blastp
545	poplar\|gb157.2\|AI165418_T1	poplar	1892	33	85	97.9166667	99.3031359	blastp
546	poplar\|gb157.2\|BU817536_T3	poplar	1893	33	82	88.1944444	92.0863309	blastp
547	poplar\|gb157.2\|BU881784_T1	poplar	1894	33	83	98.2638889	99.3031359	blastp
548	potato\|gb157.2\|BE923816_T1	potato	1895	33	85	98.2638889	99.3006993	blastp
549	potato\|gb157.2\|CK260061_T1	potato	1896	33	91	62.5	98.9010989	blastp
550	potato\|gb157.2\|BE924585_T1	potato	1897	33	86	98.2638889	98.9473684	blastp
551	potato\|gb157.2\|BF153976_T1	potato	1898	33	86	61.1111111	100	blastp
552	potato\|gb157.2\|AJ487323_T1	potato	1899	33	97	100	100	blastp
553	potato\|gb157.2\|BF154021_T1	potato	1900	33	92	99.3055556	100	blastp
554	potato\|gb157.2\|BG599633_T1	potato	1901	33	95	98.9583333	99.3031359	blastp
555	potato\|gb157.2\|BE922307_T1	potato	1902	33	85	98.2638889	99.3006993	blastp
556	radish\|gb164\|EV536875_T1	radish	1903	33	86	98.2638889	98.951049	blastp
557	radish\|gb164\|EW726189_T1	radish	1904	33	83	60.0694444	96.1111111	blastp
558	radish\|gb164\|EX756217_T1	radish	1905	33	86	98.2638889	98.951049	blastp
559	radish\|gb164\|AB030696_T1	radish	1906	33	86	98.2638889	98.951049	blastp
560	radish\|gb164\|AB030695_T1	radish	1907	33	87	98.2638889	98.951049	blastp
561	radish\|gb164\|AB012044_T1	radish	1908	33	85	98.2638889	98.951049	blastp
562	radish\|gb164\|EV567230_T1	radish	1909	33	87	98.2638889	98.9547038	blastp
563	radish\|gb164\|EY936735_T1	radish	1910	33	86	98.2638889	98.951049	blastp
564	rice\|gb157.2\|U37951_T1	rice	1911	33	86	98.2638889	98.9619377	blastp
565	rice\|gb157.2\|U40140_T1	rice	1912	33	86	98.2638889	98.9583333	blastp
566	rice\|gb157.2\|BE039992_T1	rice	1913	33	82	98.2638889	98.9583333	blastp
567	rice\|gb157.2\|U37951_T2	rice	1914	33	86	73.6111111	99.0654206	blastp
568	rose\|gb157.2\|BQ104887_T1	rose	1915	33	83	97.5694444	98.6206897	blastp
569	rose\|gb157.2\|BQ103877_T1	rose	1916	33	85	98.2638889	98.9547038	blastp
570	rose\|gb157.2\|EC586734_T1	rose	1917	33	80	62.1527778	99.4444444	blastp
571	rye\|gb164\|BE586240_T1	rye	1918	33	83	98.2638889	98.9726027	blastp
572	safflower\|gb162\|	safflower	1919	33	86	89.5833333	94.5454545	blastp
	EL407054_T1
573	safflower\|gb162\|	safflower	1920	33	85	98.2638889	92.5081433	blastp
	EL400504_T1
574	safflower\|gb162\|	safflower	1921	33	83	84.375	99.5934959	blastp
	EL400004_T1
575	sesame\|gb157.2\|	sesame	1922	33	91	57.2916667	100	blastp
	BU668587_T1
576	sesame\|gb157.2\|	sesame	1923	33	87	55.2083333	99.375	blastp
	BU669929_T1
577	sorghum\|gb161.xeno\|	sorghum	1924	33	85	98.2638889	98.9583333	blastp
	AI372377_T1
578	sorghum\|gb161.xeno\|	sorghum	1925	33	82	98.2638889	98.9655172	blastp
	AI861086_T1
579	sorghum\|gb161.xeno\|	sorghum	1926	33	86	98.2638889	98.9619377	blastp
	SBU87981_T1
580	soybean\|gb166\|CD401115_T1	soybean	1927	33	82	98.2638889	99.3031359	blastp
581	soybean\|gb166\|AW348556_T1	soybean	1928	33	84	98.6111111	99.6539792	blastp
582	soybean\|gb166\|BE352670_T1	soybean	1929	33	86	98.2638889	98.9547038	blastp
583	soybean\|gb166\|BI967765_T1	soybean	1930	33	86	98.2638889	98.9619377	blastp
584	soybean\|gb166\|BE661219_T1	soybean	1931	33	82	98.2638889	99.3079585	blastp
585	soybean\|gb166\|BE352747_T5	soybean	1932	33	81	88.1944444	98.4732824	blastp
586	soybean\|gb166\|CD416359_T1	soybean	1933	33	85	97.5694444	98.6013986	blastp
587	soybean\|gb166\|AW350352_T1	soybean	1934	33	84	97.5694444	98.6013986	blastp
588	soybean\|gb166\|BE352747_T1	soybean	1935	33	82	98.2638889	99.3079585	blastp
589	soybean\|gb166\|BE820629_T1	soybean	1936	33	85	98.2638889	99.2957746	blastp
590	spikemoss\|gb165\|	spikemoss	1937	33	82	51.0416667	99.3243243	blastp
	DN838148_T4
591	spruce\|gb162\|CO224550_T1	spruce	1938	33	80	96.875	98.9473684	blastp
592	spruce\|gb162\|CO216100_T1	spruce	1939	33	86	98.2638889	98.9726027	blastp
593	spruce\|gb162\|CO216028_T1	spruce	1940	33	85	98.2638889	98.9583333	blastp
594	spurge\|gb161\|BG354070_T1	spurge	1941	33	87	97.9166667	98.2638889	blastp
595	strawberry\|gb164\|	strawberry	1942	33	82	98.2638889	99.3103448	blastp
	CO378647_T1
596	strawberry\|gb164\|	strawberry	1943	33	84	98.2638889	98.9547038	blastp
	CX661400_T1
597	strawberry\|gb164\|	strawberry	1944	33	83	98.2638889	98.951049	blastp
	DV438296_T1
598	sugarcane\|gb157.2\|	sugarcane	1945	33	80	60.0694444	88.8888889	blastp
	CA264801_T1
599	sugarcane\|gb157.2\|	sugarcane	1946	33	85	98.2638889	98.9583333	blastp
	CA086058_T1
600	sugarcane\|gb157.2\|	sugarcane	1947	33	84	97.2222222	98.2638889	blastp
	CA071197_T1
601	sugarcane\|gb157.2\|	sugarcane	1948	33	84	91.6666667	99.2537313	blastp
	BQ530399_T1
602	sugarcane\|gb157.2\|	sugarcane	1949	33	82	74.3055556	99.5391705	blastp
	CA085969_T1
603	sugarcane\|gb157.2\|	sugarcane	1950	33	84	71.1805556	93.6936937	blastp
	CA074778_T1
604	sugarcane\|gb157.2\|	sugarcane	1951	33	81	62.1527778	99.4505495	blastp
	CA130651_T1
605	sugarcane\|gb157.2\|	sugarcane	1952	33	88	53.4722222	98.7179487	blastp
	AA525652_T1
606	sugarcane\|gb157.2\|	sugarcane	1953	33	86	98.2638889	98.9619377	blastp
	BQ536359_T1
607	sunflower\|gb162\|	sunflower	1954	33	86	98.2638889	98.9547038	blastp
	DY909123_T1
608	sunflower\|gb162\|	sunflower	1955	33	85	98.2638889	98.9547038	blastp
	CD846367_T1
609	sunflower\|gb162\|	sunflower	1956	33	86	98.2638889	98.9547038	blastp
	CD846084_T1
610	sunflower\|gb162\|	sunflower	1957	33	84	72.2222222	100	blastp
	DY915760_T1
611	sunflower\|gb162\|	sunflower	1958	33	83	73.6111111	99.5327103	blastp
	CF080940_T1
612	sunflower\|gb162\|	sunflower	1959	33	83	96.875	97.5694444	blastp
	CF087907_T1
613	sunflower\|gb162\|	sunflower	1960	33	84	98.2638889	98.6111111	blastp
	CX946986_T1
614	sunflower\|gb162\|	sunflower	1961	33	87	73.6111111	99.0697674	blastp
	DY918780_T1
615	switchgrass\|gb165\|	switchgrass	1962	33	86	98.2638889	98.9583333	blastp
	FE619753_T1
616	switchgrass\|gb165\|	switchgrass	1963	33	85	98.2638889	98.9583333	blastp
	DN142591_T1
617	switchgrass\|gb165\|	switchgrass	1964	33	85	98.2638889	98.9619377	blastp
	DN141716_T1
618	switchgrass\|gb165\|	switchgrass	1965	33	86	98.2638889	98.9583333	blastp
	DN141343_T1
619	switchgrass\|gb165\|	switchgrass	1966	33	85	98.2638889	98.9619377	blastp
	DN142037_T1
620	thellungiella\|gb157.2\|	thellungiella	1967	33	85	98.2638889	98.951049	blastp
	DN774595_T1
621	thellungiella\|gb157.2\|	thellungiella	1968	33	86	98.2638889	98.951049	blastp
	BM986095_T1
622	thellungiella\|gb157.2\|	thellungiella	1969	33	85	72.5694444	99.5238095	blastp
	BI698563_T1
623	tobacco\|gb162\|EB426225_T1	tobacco	1970	33	87	98.2638889	98.2578397	blastp
624	tobacco\|gb162\|CK720591_T1	tobacco	1971	33	95	99.3055556	99.6515679	blastp
625	tobacco\|gb162\|CK720595_T1	tobacco	1972	33	96	73.6111111	99.0654206	blastp
626	tobacco\|gb162\|EB427872_T1	tobacco	1973	33	88	98.2638889	99.2982456	blastp
627	tobacco\|gb162\|CK720593_T1	tobacco	1974	33	87	97.9166667	98.9473684	blastp
628	tobacco\|gb162\|AF024511_T1	tobacco	1975	33	95	99.3055556	99.6515679	blastp
629	tobacco\|gb162\|CK720596_T1	tobacco	1976	33	96	98.9583333	99.3031359	blastp
630	tobacco\|gb162\|NTU62280_T1	tobacco	1977	33	96	98.9583333	99.3031359	blastp
631	tomato\|gb164\|AW622243_T1	tomato	1978	33	94	98.9583333	99.3031359	blastp
632	tomato\|gb164\|BG123213_T1	tomato	1979	33	94	99.3055556	100	blastp
633	tomato\|gb164\|AI637363_T1	tomato	1980	33	87	98.2638889	98.9473684	blastp
634	tomato\|gb164\|BG123955_T1	tomato	1981	33	85	98.2638889	99.3006993	blastp
635	tomato\|gb164\|BP876517_T1	tomato	1982	33	80	55.9027778	86.8705036	tblastn
636	triphysaria\|gb164\|	triphysaria	1983	33	86	73.6111111	99.5305164	blastp
	BM357654_T1
637	triphysaria\|gb164\|	triphysaria	1984	33	90	65.625	99.4736842	blastp
	EY141207_T1
638	triphysaria\|gb164\|	triphysaria	1985	33	87	69.0972222	100	blastp
	DR174621_T1
639	triphysaria\|gb164\|	triphysaria	1986	33	86	88.5416667	99.6108949	blastp
	BM356761_T1
640	triphysaria\|gb164\|	triphysaria	1987	33	87	98.2638889	99.6466431	blastp
	DR169763_T1
641	triphysaria\|gb164\|	triphysaria	1988	33	86	96.5277778	99.6428571	blastp
	BM356902_T1
642	triphysaria\|gb164\|	triphysaria	1989	33	86	92.7083333	97.4545455	blastp
	DR174271_T1
643	triphysaria\|gb164\|	triphysaria	1990	33	88	100	99.6539792	blastp
	DR171777_T1
644	wheat\|gb164\|BE406715_T1	wheat	1991	33	83	98.2638889	98.9726027	blastp
645	wheat\|gb164\|BQ838456_T1	wheat	1992	33	83	98.2638889	98.9726027	blastp
646	wheat\|gb164\|BE426386_T1	wheat	1993	33	82	98.2638889	98.9726027	blastp
647	wheat\|gb164\|BE403388_T1	wheat	1994	33	88	51.0416667	98.6577181	blastp
648	wheat\|gb164\|BE403307_T1	wheat	1995	33	83	98.2638889	98.9726027	blastp
649	wheat\|gb164\|BE498268_T1	wheat	1996	33	81	60.4166667	96.7213115	blastp
650	wheat\|gb164\|AL828763_T1	wheat	1997	33	84	84.7222222	96.4705882	blastp
651	wheat\|gb164\|AF139816_T1	wheat	1998	33	83	98.2638889	98.9726027	blastp
652	wheat\|gb164\|BE216990_T1	wheat	1999	33	86	98.2638889	98.9583333	blastp
653	wheat\|gb164\|BE403886_T1	wheat	2000	33	85	99.3055556	99.6551724	blastp
654	wheat\|gb164\|BE430165_T1	wheat	2001	33	85	99.3055556	99.6551724	blastp
655	wheat\|gb164\|CA484202_T1	wheat	2002	33	87	56.9444444	97.6190476	blastp
656	wheat\|gb164\|BE404199_T1	wheat	2003	33	86	98.2638889	98.9583333	blastp
657	wheat\|gb164\|BE406086_T1	wheat	2004	33	83	98.2638889	98.9726027	blastp
658	wheat\|gb164\|BF293776_T1	wheat	2005	33	83	98.2638889	98.9655172	blastp
659	wheat\|gb164\|CK193386_T1	wheat	2006	33	81	97.2222222	96.6216216	blastp
660	castorbean\|gb160\|	castorbean	2007	34	80	99.1902834	98.7854251	blastp
	AJ605572_T1
661	citrus\|gb157.2\|CK740163_T1	citrus	2008	34	80	99.1902834	98.7854251	blastp
662	coffea\|gb157.2\|DV664793_T1	coffea	2009	34	80	100	100	blastp
663	lettuce\|gb157.2\|	lettuce	2010	34	80	99.1902834	99.5934959	blastp
	DW074942_T1
664	pepper\|gb157.2\|	pepper	2011	34	95	76.1133603	100	blastp
	BM063938_T1
665	periwinkle\|gb164\|	periwinkle	2012	34	81	99.1902834	98.7903226	blastp
	EG558295_T1
666	potato\|gb157.2\|BM112462_T1	potato	2013	34	98	94.7368421	99.5744681	blastp
667	tobacco\|gb162\|AJ237751_T1	tobacco	2014	34	95	100	100	blastp
668	tobacco\|gb162\|EB425012_T1	tobacco	2015	34	93	100	100	blastp
669	nicotiana_benthamiana\|	nicotiana_benthamiana	2016	35	86	74.617737	97.5903614	blastp
	gb162\|CK284579_T1
670	potato\|gb157.2\|BG594926_T1	potato	2017	35	96	100	96.4497041	blastp
671	tomato\|gb164\|DB679435_T1	tomato	2018	35	90	76.146789	100	blastp
672	tobacco\|gb162\|CK720588_T1	tobacco	2019	36	81	53.5580524	100	blastp
673	apple\|gb157.3\|CN494715_T1	apple	2020	37	84	85.7142857	94.7368421	blastp
674	apple\|gb157.3\|CO066689_T1	apple	2021	37	81	94.2857143	76.1538462	blastp
675	castorbean\|gb160\|	castorbean	2022	37	90	95.2380952	36.900369	blastp
	MDL30026M001488_T1
676	citrus\|gb157.2\|CX300349_T1	citrus	2023	37	83	99.047619	72.7272727	blastp
677	coffea\|gb157.2\|DV663640_T1	coffea	2024	37	80	93.3333333	34.5070423	blastp
678	cowpea\|gb166\|FF395821_T1	cowpea	2025	37	82	93.3333333	95.1456311	blastp
679	cowpea\|gb166\|FF395821_T2	cowpea	2026	37	82	94.2857143	13.2450331	tblastn
680	ipomoea\|gb157.2\|	ipomoea	2027	37	87	100	59.3220339	blastp
	CJ769054_T1
681	lettuce\|gb157.2\|	lettuce	2028	37	83	98.0952381	36.5248227	blastp
	CV699989_T1
682	medicago\|gb157.2\|	medicago	2029	37	80	95.2380952	37.1747212	blastp
	AW208262_T1
683	melon\|gb165\|AM727408_T1	melon	2030	37	82	97.1428571	36.9565217	blastp
684	poplar\|gb157.2\|CN517706_T1	poplar	2031	37	84	96.1904762	36.5942029	blastp
685	poplar\|gb157.2\|BU813630_T1	poplar	2032	37	84	99.047619	37.6811594	blastp
686	potato\|gb157.2\|DN587628_T1	potato	2033	37	90	96.1904762	93.5185185	blastp
687	rice\|gb157.2\|BI805522_T1	rice	2034	37	80	97.1428571	35.915493	blastp
688	soybean\|gb166\|FK397604_T1	soybean	2035	37	82	54.2857143	98.2758621	blastp
689	sunflower\|gb162\|	sunflower	2036	37	82	98.0952381	39.0151515	blastp
	DY951259_T1
690	sunflower\|gb162\|	sunflower	2037	37	83	98.0952381	37.3188406	blastp
	DY942645_T1
691	triphysaria\|gb164\|	triphysaria	2038	37	85	99.047619	37.6811594	blastp
	EY130232_T1
692	arabidopsis\|gb165\|	arabidopsis	2039	38	80	97.6271186	96.0526316	blastp
	AT4G10380_T1
693	artemisia\|gb164\|	artemisia	2040	38	87	55.5932203	100	blastp
	EY089420_T1
694	artemisia\|gb164\|	artemisia	2041	38	89	51.8644068	100	blastp
	EY113317_T1
695	b_oleracea\|gb161\|	b_oleracea	2042	38	81	93.8983051	95.862069	blastp
	AM391026_T1
696	b_rapa\|gb162\|CV545128_T1	b_rapa	2043	38	81	97.6271186	96.013289	blastp
697	canola\|gb161\|ES903871_T1	canola	2044	38	85	52.2033898	100	blastp
698	cassava\|gb164\|CK641734_T1	cassava	2045	38	89	93.8983051	98.9247312	blastp
699	castorbean\|gb160\|	castorbean	2046	38	85	100	100	blastp
	EG668085_T1
700	castorbean\|gb160\|	castorbean	2047	38	90	62.0338983	100	blastp
	EG668085_T2
701	centaurea\|gb161\|	centaurea	2048	38	83	69.1525424	98.0487805	blastp
	EH739099_T1
702	centaurea\|gb161\|	centaurea	2049	38	89	80.6779661	95.9677419	blastp
	EH710762_T1
703	citrus\|gb157.2\|CX299695_T1	citrus	2050	38	98	62.0338983	100	blastp
704	citrus\|gb157.2\|CO912981_T1	citrus	2051	38	80	100	100	blastp
705	citrus\|gb157.2\|CO912981_T2	citrus	2052	38	81	78.3050847	95.5465587	blastp
706	cotton\|gb164\|CO071578_T1	cotton	2053	38	90	77.9661017	95.4356846	blastp
707	cotton\|gb164\|BE052767_T1	cotton	2054	38	88	86.440678	100	blastp
708	grape\|gb160\|CB350030_T1	grape	2055	38	86	100	100	blastp
709	lettuce\|gb157.2\|	lettuce	2056	38	86	97.6271186	96.6555184	blastp
	DW123895_T1
710	melon\|gb165\|AM726471_T1	melon	2057	38	80	78.6440678	92.8	blastp
711	nicotiana_benthamiana\|	nicotiana_benthamiana	2058	38	94	100	100	blastp
	gb162\|CK281387_T1
712	onion\|gb162\|CF436356_T1	onion	2059	38	83	86.1016949	98.0988593	blastp
713	poplar\|gb157.2\|BU895174_T1	poplar	2060	38	84	97.6271186	96.3333333	blastp
714	poplar\|gb157.2\|BI126692_T1	poplar	2061	38	85	100	100	blastp
715	radish\|gb164\|EX772276_T1	radish	2062	38	87	62.0338983	100	blastp
716	safflower\|gb162\|	safflower	2063	38	87	55.5932203	94.2528736	blastp
	EL376221_T1
717	soybean\|gb166\|AW351195_T1	soybean	2064	38	83	57.2881356	100	blastp
718	spurge\|gb161\|DV120704_T1	spurge	2065	38	81	66.1016949	100	blastp
719	strawberry\|gb164\|	strawberry	2066	38	87	73.559322	100	blastp
	EX663538_T1
720	sunflower\|gb162\|	sunflower	2067	38	83	69.8305085	95.0636943	tblastn
	BQ915292_T1
721	tobacco\|gb162\|EB426773_T1	tobacco	2068	38	94	100	100	blastp
722	triphysaria\|gb164\|	triphysaria	2069	38	90	67.7966102	100	blastp
	EY008469_T1
723	pepper\|gb157.2\|	pepper	2070	39	91	60	100	blastp
	BM066463_T1
724	tobacco\|gb162\|EB445778_T1	tobacco	2071	39	88	85.8333333	100	blastp
725	pepper\|gb157.2\|	pepper	2072	40	82	64.4628099	100	blastp
	CA515996_T1
726	potato\|gb157.2\|BE341068_T1	potato	2073	40	92	100	100	blastp
727	potato\|gb157.2\|BG887984_T1	potato	2074	41	100	79.4238683	91.4691943	blastp
728	apple\|gb157.3\|CN898142_T1	apple	2075	42	86	70.9677419	98.0582524	blastp
729	apple\|gb157.3\|CN492544_T1	apple	2076	42	82	99.6415771	98.9399293	blastp
730	apple\|gb157.3\|CN495819_T1	apple	2077	42	84	99.6415771	98.9547038	blastp
731	apple\|gb157.3\|CN869175_T1	apple	2078	42	86	100	100	blastp
732	apple\|gb157.3\|CN488973_T1	apple	2079	42	87	100	100	blastp
733	apricot\|gb157.2\|	apricot	2080	42	88	69.8924731	98.4848485	blastp
	CB820380_T1
734	aquilegia\|gb157.3\|	aquilegia	2081	42	85	94.9820789	100	blastp
	DR921860_T1
735	arabidopsis\|gb165\|	arabidopsis	2082	42	80	100	99.2982456	blastp
	AT2G37170_T1
736	arabidopsis\|gb165\|	arabidopsis	2083	42	81	95.6989247	94.7552448	blastp
	AT3G54820_T1
737	arabidopsis\|gb165\|	arabidopsis	2084	42	81	100	99.3031359	blastp
	AT3G53420_T1
738	arabidopsis\|gb165\|	arabidopsis	2085	42	80	99.2831541	97.2508591	blastp
	AT5G60660_T1
739	artemisia\|gb164\|	artemisia	2086	42	89	79.5698925	100	blastp
	EY056827_T1
740	artemisia\|gb164\|	artemisia	2087	42	83	100	100	blastp
	EY033689_T1
741	artemisia\|gb164\|	artemisia	2088	42	81	100	100	blastp
	EY032199_T1
742	artemisia\|gb164\|	artemisia	2089	42	88	100	100	blastp
	EY042731_T1
743	artemisia\|gb164\|	artemisia	2090	42	82	99.6415771	98.9473684	blastp
	EX980079_T1
744	avocado\|gb164\|CK754546_T1	avocado	2091	42	86	51.6129032	97.2972973	blastp
745	b_juncea\|gb164\|	b_juncea	2092	42	81	100	99.2982456	blastp
	EVGN00454408761136_T1
746	b_juncea\|gb164\|	b_juncea	2093	42	81	98.9247312	97.9020979	blastp
	EVGN00454408761136_T2
747	b_juncea\|gb164\|	b_juncea	2094	42	80	100	88.7850467	blastp
	EVGN00748222952488_T2
748	b_juncea\|gb164\|	b_juncea	2095	42	84	82.078853	97.8991597	blastp
	EVGN00204411253360_T1
749	b_juncea\|gb164\|	b_juncea	2096	42	80	100	99.2982456	blastp
	EVGN00054208600715_T1
750	b_juncea\|gb164\|	b_juncea	2097	42	83	64.516129	96.2566845	blastp
	EVGN00049614332152_T1
751	b_juncea\|gb164\|	b_juncea	2098	42	85	58.4229391	100	blastp
	EVGN01023711071914_T1
752	b_juncea\|gb164\|	b_juncea	2099	42	81	100	99.3031359	blastp
	EVGN00247216171316_T1
753	b_juncea\|gb164\|	b_juncea	2100	42	88	64.1577061	100	blastp
	EVGN00778009020884_T1
754	b_juncea\|gb164\|	b_juncea	2101	42	85	53.4050179	98.6754967	blastp
	EVGN02648808940517_T1
755	b_juncea\|gb164\|	b_juncea	2102	42	82	82.7956989	100	blastp
	EVGN00316414413452_T1
756	b_juncea\|gb164\|	b_juncea	2103	42	81	100	99.3031359	blastp
	DT317706_T1
757	b_juncea\|gb164\|	b_juncea	2104	42	81	100	99.3031359	blastp
	EVGN00748222952488_T1
758	b_oleracea\|gb161\|	b_oleracea	2105	42	80	100	100	blastp
	AM386520_T1
759	b_oleracea\|gb161\|	b_oleracea	2106	42	83	57.3476703	91.954023	blastp
	AM058395_T1
760	b_oleracea\|gb161\|	b_oleracea	2107	42	81	100	99.2982456	blastp
	AM385504_T1
761	b_rapa\|gb162\|BG544498_T1	b_rapa	2108	42	81	100	100	blastp
762	b_rapa\|gb162\|BQ791962_T2	b_rapa	2109	42	81	100	99.2982456	blastp
763	b_rapa\|gb162\|BQ791962_T1	b_rapa	2110	42	81	100	99.2982456	blastp
764	b_rapa\|gb162\|EX065729_T1	b_rapa	2111	42	81	92.1146953	100	blastp
765	b_rapa\|gb162\|CA992278_T1	b_rapa	2112	42	83	89.9641577	98.8372093	blastp
766	b_rapa\|gb162\|CO749284_T1	b_rapa	2113	42	81	100	99.2982456	blastp
767	barley\|gb157.3\|BE412486_T1	barley	2114	42	81	100	100	blastp
768	bean\|gb164\|CB542746_T1	bean	2115	42	80	100	100	blastp
769	bean\|gb164\|CB280567_T1	bean	2116	42	86	99.6415771	98.9473684	blastp
770	bean\|gb164\|BQ481649_T1	bean	2117	42	82	100	100	blastp
771	bean\|gb164\|CV532291_T1	bean	2118	42	86	99.6415771	98.9547038	blastp
772	brachypodium\|gb161.xeno\|	brachypodium	2119	42	80	94.9820789	100	blastp
	BE416137_T1
773	brachypodium\|gb161.xeno\|	brachypodium	2120	42	80	100	100	blastp
	AF139814_T1
774	canola\|gb161\|CN729066_T1	canola	2121	42	81	100	99.3031359	blastp
775	canola\|gb161\|DQ068169_T1	canola	2122	42	81	100	99.2982456	blastp
776	canola\|gb161\|AF118382_T1	canola	2123	42	81	100	99.3031359	blastp
777	canola\|gb161\|AF118383_T1	canola	2124	42	81	100	100	blastp
778	canola\|gb161\|CD819509_T1	canola	2125	42	80	100	99.2982456	blastp
779	canola\|gb161\|EE419467_T1	canola	2126	42	81	100	100	blastp
780	canola\|gb161\|EE459735_T1	canola	2127	42	81	100	99.2982456	blastp
781	canola\|gb161\|CX192356_T1	canola	2128	42	80	100	100	blastp
782	canola\|gb161\|CN827413_T1	canola	2129	42	81	100	99.2982456	blastp
783	canola\|gb161\|EE432011_T1	canola	2130	42	81	100	99.2982456	blastp
784	cassava\|gb164\|DB922106_T1	cassava	2131	42	81	93.90681	92.5266904	blastp
785	cassava\|gb164\|CK640888_T1	cassava	2132	42	83	100	100	blastp
786	cassava\|gb164\|CK642866_T1	cassava	2133	42	84	99.6415771	98.951049	blastp
787	cassava\|gb164\|CK642551_T1	cassava	2134	42	86	100	99.3055556	blastp
788	castorbean\|gb160\|	castorbean	2135	42	86	100	99.3055556	blastp
	AJ605565_T1
789	castorbean\|gb160\|	castorbean	2136	42	87	99.6415771	98.9399293	blastp
	AJ605568_T1
790	castorbean\|gb160\|	castorbean	2137	42	85	100	100	blastp
	EE259660_T1
791	centaurea\|gb161\|	centaurea	2138	42	84	87.0967742	92.8571429	blastp
	EL933765_T1
792	centaurea\|gb161\|	centaurea	2139	42	82	75.9856631	89.2561983	blastp
	EL931761_T1
793	cichorium\|gb161\|	cichorium	2140	42	86	100	100	blastp
	DT212328_T1
794	citrus\|gb157.2\|BQ625054_T1	citrus	2141	42	83	100	100	blastp
795	citrus\|gb157.2\|CF509045_T1	citrus	2142	42	86	100	99.3031359	blastp
796	citrus\|gb157.2\|CB291797_T1	citrus	2143	42	80	100	84.0707965	blastp
797	citrus\|gb157.2\|BQ623325_T1	citrus	2144	42	86	100	99.3031359	blastp
798	citrus\|gb157.2\|BQ623742_T1	citrus	2145	42	88	94.9820789	99.2619926	blastp
799	citrus\|gb157.2\|CF417769_T1	citrus	2146	42	83	100	99.3031359	blastp
800	citrus\|gb157.2\|BQ623128_T1	citrus	2147	42	88	68.4587814	98.9637306	blastp
801	citrus\|gb157.2\|CB293000_T1	citrus	2148	42	82	93.1899642	98.8847584	blastp
802	citrus\|gb157.2\|BQ622991_T1	citrus	2149	42	84	96.4157706	98.5559567	blastp
803	citrus\|gb157.2\|CF503882_T1	citrus	2150	42	83	100	100	blastp
804	cotton\|gb164\|BE052942_T1	cotton	2151	42	84	99.2831541	98.5815603	blastp
805	cotton\|gb164\|AF064467_T1	cotton	2152	42	80	100	100	blastp
806	cotton\|gb164\|BG443494_T1	cotton	2153	42	85	100	99.2982456	blastp
807	cotton\|gb164\|CO086106_T1	cotton	2154	42	88	70.2508961	100	blastp
808	cotton\|gb164\|BQ406033_T1	cotton	2155	42	82	100	99.2982456	blastp
809	cotton\|gb164\|CO109551_T1	cotton	2156	42	82	100	100	blastp
810	cotton\|gb164\|DV437970_T1	cotton	2157	42	80	64.1577061	95.3608247	blastp
811	cotton\|gb164\|AI725803_T1	cotton	2158	42	84	100	99.2982456	blastp
812	cotton\|gb164\|CD486305_T1	cotton	2159	42	81	98.9247312	94.0199336	blastp
813	cowpea\|gb166\|ES884222_T2	cowpea	2160	42	85	86.3799283	93.5606061	blastp
814	cowpea\|gb166\|ES884222_T1	cowpea	2161	42	86	99.6415771	98.9547038	blastp
815	cowpea\|gb166\|FF538675_T1	cowpea	2162	42	89	52.688172	98	blastp
816	cowpea\|gb166\|FC458151_T1	cowpea	2163	42	80	100	100	blastp
817	cowpea\|gb166\|FC458381_T1	cowpea	2164	42	85	99.6415771	98.9473684	blastp
818	cryptomeria\|gb166\|	cryptomeria	2165	42	83	95.6989247	93.639576	blastp
	AU036821_T1
819	cryptomeria\|gb166\|	cryptomeria	2166	42	81	53.046595	98.013245	blastp
	BW995927_T1
820	dandelion\|gb161\|	dandelion	2167	42	87	93.90681	88.9632107	blastp
	DY827637_T1
821	dandelion\|gb161\|	dandelion	2168	42	85	93.5483871	93.2862191	blastp
	DY814583_T1
822	dandelion\|gb161\|	dandelion	2169	42	85	100	100	blastp
	DY828216_T1
823	dandelion\|gb161\|	dandelion	2170	42	80	85.3046595	98.7654321	blastp
	DY818322_T1
824	dandelion\|gb161\|	dandelion	2171	42	82	98.9247312	98.245614	blastp
	DY805523_T1
825	fescue\|gb161\|DT675934_T1	fescue	2172	42	82	61.2903226	93.9226519	blastp
826	ginger\|gb164\|DY345807_T1	ginger	2173	42	81	100	100	blastp
827	ginger\|gb164\|DY373920_T1	ginger	2174	42	88	69.5340502	100	blastp
828	grape\|gb160\|BQ792080_T1	grape	2175	42	81	100	99.3031359	blastp
829	grape\|gb160\|CB973593_T1	grape	2176	42	84	100	100	blastp
830	grape\|gb160\|BM437196_T1	grape	2177	42	84	100	99.2957746	blastp
831	iceplant\|gb164\|CIPMIPC_T1	iceplant	2178	42	83	100	100	blastp
832	iceplant\|gb164\|BE035661_T1	iceplant	2179	42	82	99.6415771	98.6254296	blastp
833	ipomoea\|gb157.2\|	ipomoea	2180	42	80	99.6415771	100	blastp
	BM878800_T1
834	ipomoea\|gb157.2\|	ipomoea	2181	42	90	86.7383513	93.2330827	blastp
	AU224434_T2
835	ipomoea\|gb157.2\|	ipomoea	2182	42	82	100	99.2957746	blastp
	BJ553793_T1
836	ipomoea\|gb157.2\|	ipomoea	2183	42	89	100	100	blastp
	AU224434_T1
837	lettuce\|gb157.2\|	lettuce	2184	42	85	99.2831541	98.5964912	blastp
	DW115660_T1
838	lettuce\|gb157.2\|	lettuce	2185	42	80	97.8494624	95.7894737	blastp
	DW113963_T1
839	lettuce\|gb157.2\|	lettuce	2186	42	84	99.6415771	98.9399293	blastp
	DW110249_T1
840	lettuce\|gb157.2\|	lettuce	2187	42	86	100	100	blastp
	DW051453_T1
841	lettuce\|gb157.2\|	lettuce	2188	42	84	100	100	blastp
	DW076507_T1
842	lettuce\|gb157.2\|	lettuce	2189	42	84	100	100	blastp
	DW127617_T1
843	lettuce\|gb157.2\|	lettuce	2190	42	80	100	100	blastp
	AJ937963_T1
844	lettuce\|gb157.2\|	lettuce	2191	42	83	92.8315412	98.5018727	blastp
	DW049421_T1
845	lettuce\|gb157.2\|	lettuce	2192	42	87	99.2831541	98.9473684	blastp
	DW114305_T1
846	lettuce\|gb157.2\|	lettuce	2193	42	80	100	100	blastp
	DW053722_T1
847	lettuce\|gb157.2\|	lettuce	2194	42	81	100	100	blastp
	DW146178_T1
848	lettuce\|gb157.2\|	lettuce	2195	42	85	99.2831541	98.5964912	blastp
	DW077710_T1
849	lettuce\|gb157.2\|	lettuce	2196	42	84	100	100	blastp
	DW070566_T1
850	lettuce\|gb157.2\|	lettuce	2197	42	85	94.9820789	100	blastp
	DW093078_T1
851	lettuce\|gb157.2\|	lettuce	2198	42	84	97.4910394	96.1672474	blastp
	DW074446_T1
852	lettuce\|gb157.2\|	lettuce	2199	42	83	99.6415771	98.9399293	blastp
	DW153482_T1
853	lettuce\|gb157.2\|	lettuce	2200	42	86	100	100	blastp
	DW080306_T1
854	lettuce\|gb157.2\|	lettuce	2201	42	84	99.6415771	98.9399293	blastp
	DW043674_T1
855	lettuce\|gb157.2\|	lettuce	2202	42	83	97.1326165	98.5559567	blastp
	DW095979_T1
856	lettuce\|gb157.2\|	lettuce	2203	42	84	100	100	blastp
	DW077206_T1
857	lettuce\|gb157.2\|	lettuce	2204	42	86	98.9247312	99.6453901	blastp
	DW047573_T1
858	lettuce\|gb157.2\|	lettuce	2205	42	87	100	100	blastp
	DW096304_T1
859	lettuce\|gb157.2\|	lettuce	2206	42	81	100	100	blastp
	DW075191_T1
860	lotus\|gb157.2\|AI967757_T1	lotus	2207	42	85	99.6415771	98.9547038	blastp
861	lotus\|gb157.2\|AI967387_T2	lotus	2208	42	80	99.6415771	99.6515679	blastp
862	lotus\|gb157.2\|BG662315_T1	lotus	2209	42	82	99.6415771	98.9619377	blastp
863	maize\|gb164\|BE552783_T1	maize	2210	42	80	97.4910394	95.890411	blastp
864	maize\|gb164\|AI622334_T1	maize	2211	42	83	97.4910394	95.862069	blastp
865	maize\|gb164\|AI855280_T1	maize	2212	42	81	96.7741935	95.1557093	blastp
866	medicago\|gb157.2\|	medicago	2213	42	80	100	99.3031359	blastp
	AW981259_T1
867	medicago\|gb157.2\|	medicago	2214	42	83	99.6415771	98.9547038	blastp
	AA660788_T1
868	melon\|gb165\|DV631824_T1	melon	2215	42	84	100	100	blastp
869	melon\|gb165\|DV633977_T1	melon	2216	42	83	64.516129	100	blastp
870	melon\|gb165\|AM720039_T1	melon	2217	42	81	86.0215054	100	blastp
871	nicotiana_benthamiana\|	nicotiana_benthamiana	2218	42	80	100	100	blastp
	gb162\|CN743200_T1
872	nicotiana_benthamiana\|	nicotiana_benthamiana	2219	42	80	98.2078853	96.8641115	blastp
	gb162\|CK294539_T1
873	onion\|gb162\|AF255795_T1	onion	2220	42	82	97.1326165	95.532646	blastp
874	onion\|gb162\|CF434704_T1	onion	2221	42	80	99.2831541	99.2957746	blastp
875	papaya\|gb165\|EL784273_T1	papaya	2222	42	82	82.078853	97.5206612	blastp
876	papaya\|gb165\|AM904340_T1	papaya	2223	42	84	100	99.2882562	blastp
877	peach\|gb157.2\|AF367458_T1	peach	2224	42	84	87.0967742	100	blastp
878	peach\|gb157.2\|BU040116_T1	peach	2225	42	83	99.6415771	98.9547038	blastp
879	peach\|gb157.2\|AF367460_T1	peach	2226	42	86	87.0967742	100	blastp
880	peanut\|gb161\|CD037924_T1	peanut	2227	42	86	99.6415771	98.9547038	blastp
881	peanut\|gb161\|CD038296_T1	peanut	2228	42	85	99.6415771	98.9547038	blastp
882	peanut\|gb161\|CD037924_T2	peanut	2229	42	86	99.6415771	98.9547038	blastp
883	peanut\|gb161\|CD037884_T1	peanut	2230	42	88	51.2544803	97.9452055	blastp
884	pepper\|gb157.2\|	pepper	2231	42	96	100	100	blastp
	BM061612_T1
885	pepper\|gb157.2\|	pepper	2232	42	97	100	100	blastp
	BM061611_T1
886	pepper\|gb157.2\|	pepper	2233	42	81	92.4731183	97.3977695	blastp
	BM061005_T1
887	petunia\|gb157.2\|	petunia	2234	42	83	97.4910394	98.2332155	blastp
	AF452014_T1
888	petunia\|gb157.2\|	petunia	2235	42	95	53.046595	93.6708861	blastp
	CV295523_T1
889	petunia\|gb157.2\|	petunia	2236	42	83	55.5555556	100	blastp
	CV293001_T1
890	pine\|gb157.2\|AI813221_T1	pine	2237	42	81	96.0573477	94.3262411	blastp
891	pine\|gb157.2\|CA844411_T1	pine	2238	42	81	56.6308244	98.75	blastp
892	poplar\|gb157.2\|BI130501_T3	poplar	2239	42	85	100	99.2982456	blastp
893	poplar\|gb157.2\|AI165755_T1	poplar	2240	42	86	99.6415771	98.9473684	blastp
894	poplar\|gb157.2\|BU835712_T1	poplar	2241	42	85	99.6415771	98.9473684	blastp
895	poplar\|gb157.2\|BI130501_T1	poplar	2242	42	85	100	99.2982456	blastp
896	poplar\|gb157.2\|BI130501_T4	poplar	2243	42	85	100	99.2982456	blastp
897	poplar\|gb157.2\|AI162288_T1	poplar	2244	42	86	99.2831541	98.5964912	blastp
898	poplar\|gb157.2\|AJ534524_T1	poplar	2245	42	84	100	100	blastp
899	potato\|gb157.2\|BG096672_T1	potato	2246	42	96	100	100	blastp
900	potato\|gb157.2\|BM109370_T1	potato	2247	42	80	98.2078853	96.8641115	blastp
901	potato\|gb157.2\|BG589618_T1	potato	2248	42	80	98.2078853	96.8641115	blastp
902	potato\|gb157.2\|CK261080_T1	potato	2249	42	83	70.2508961	100	blastp
903	potato\|gb157.2\|BG098124_T1	potato	2250	42	97	99.2831541	100	blastp
904	potato\|gb157.2\|BI406400_T1	potato	2251	42	80	98.2078853	96.8641115	blastp
905	potato\|gb157.2\|BE920139_T1	potato	2252	42	90	100	100	blastp
906	potato\|gb157.2\|BM112017_T1	potato	2253	42	80	98.2078853	96.8641115	blastp
907	potato\|gb157.2\|BE921679_T1	potato	2254	42	96	100	100	blastp
908	potato\|gb157.2\|BG600158_T1	potato	2255	42	80	98.2078853	96.8641115	blastp
909	potato\|gb157.2\|BI406047_T1	potato	2256	42	80	100	100	blastp
910	potato\|gb157.2\|CK719282_T1	potato	2257	42	80	98.2078853	96.8641115	blastp
911	radish\|gb164\|EV545956_T1	radish	2258	42	81	100	99.2982456	blastp
912	radish\|gb164\|EX904869_T1	radish	2259	42	80	100	100	blastp
913	radish\|gb164\|EV545247_T1	radish	2260	42	80	100	100	blastp
914	radish\|gb164\|EX756889_T1	radish	2261	42	80	100	100	blastp
915	radish\|gb164\|EV539533_T1	radish	2262	42	81	100	99.3031359	blastp
916	radish\|gb164\|EV546186_T1	radish	2263	42	82	55.5555556	95.6790123	blastp
917	radish\|gb164\|AB012045_T1	radish	2264	42	81	100	99.3031359	blastp
918	radish\|gb164\|EV573001_T1	radish	2265	42	83	62.0071685	98.8571429	blastp
919	radish\|gb164\|AB030698_T1	radish	2266	42	81	100	100	blastp
920	radish\|gb164\|EX749049_T1	radish	2267	42	83	78.4946237	100	blastp
921	radish\|gb164\|AB030697_T1	radish	2268	42	80	100	100	blastp
922	radish\|gb164\|EW735060_T1	radish	2269	42	80	96.4157706	95.4545455	blastp
923	radish\|gb164\|FD936119_T1	radish	2270	42	83	50.5376344	100	blastp
924	radish\|gb164\|EV543747_T1	radish	2271	42	81	100	99.3031359	blastp
925	rice\|gb157.2\|BE040651_T2	rice	2272	42	80	86.7383513	94.3820225	blastp
926	rice\|gb157.2\|BE040651_T1	rice	2273	42	80	100	100	blastp
927	rice\|gb157.2\|AA754435_T5	rice	2274	42	80	85.6630824	87.7192982	blastp
928	rice\|gb157.2\|AA754435_T1	rice	2275	42	81	100	100	blastp
929	rose\|gb157.2\|BI977420_T1	rose	2276	42	84	69.8924731	91.627907	blastp
930	rose\|gb157.2\|BI977750_T1	rose	2277	42	84	77.7777778	100	blastp
931	rose\|gb157.2\|BI978110_T1	rose	2278	42	83	64.516129	98.3783784	blastp
932	safflower\|gb162\|	safflower	2279	42	81	98.5663082	100	blastp
	EL406648_T1
933	safflower\|gb162\|	safflower	2280	42	86	92.4731183	97.761194	blastp
	EL411110_T1
934	safflower\|gb162\|	safflower	2281	42	85	100	100	blastp
	EL401452_T1
935	safflower\|gb162\|	safflower	2282	42	83	97.1326165	100	blastp
	EL402424_T1
936	safflower\|gb162\|	safflower	2283	42	85	86.7383513	67.9193401	tblastn
	EL400227_T1
937	sorghum\|gb161.xeno\|	sorghum	2284	42	83	97.4910394	95.862069	blastp
	AI622334_T1
938	soybean\|gb166\|CD393286_T1	soybean	2285	42	80	99.6415771	99.6515679	blastp
939	soybean\|gb166\|GMU27347_T1	soybean	2286	42	86	99.6415771	98.9473684	blastp
940	soybean\|gb166\|AW349289_T1	soybean	2287	42	85	99.6415771	98.9473684	blastp
941	soybean\|gb166\|BE823946_T1	soybean	2288	42	87	99.6415771	98.943662	blastp
942	soybean\|gb166\|BE352729_T2	soybean	2289	42	80	56.2724014	100	blastp
943	soybean\|gb166\|AW350475_T1	soybean	2290	42	85	99.6415771	98.9547038	blastp
944	soybean\|gb166\|BI119558_T1	soybean	2291	42	80	100	100	blastp
945	soybean\|gb166\|AW349392_T1	soybean	2292	42	80	100	100	blastp
946	spruce\|gb162\|AF051202_T1	spruce	2293	42	81	96.0573477	94.3262411	blastp
947	spruce\|gb162\|CO227554_T1	spruce	2294	42	80	96.0573477	93.6619718	blastp
948	spruce\|gb162\|CO220480_T1	spruce	2295	42	80	96.4157706	94.6808511	blastp
949	spruce\|gb162\|CO217151_T1	spruce	2296	42	80	96.0573477	94.3262411	blastp
950	spurge\|gb161\|AW990929_T1	spurge	2297	42	80	59.8566308	91.5343915	blastp
951	spurge\|gb161\|BG354126_T1	spurge	2298	42	83	100	100	blastp
952	spurge\|gb161\|DV125170_T1	spurge	2299	42	87	83.5125448	99.5780591	blastp
953	strawberry\|gb164\|	strawberry	2300	42	85	99.6415771	98.9473684	blastp
	DV438166_T1
954	strawberry\|gb164\|	strawberry	2301	42	86	100	99.2957746	blastp
	EX665494_T1
955	strawberry\|gb164\|	strawberry	2302	42	85	100	100	blastp
	CO817390_T1
956	sugarcane\|gb157.2\|	sugarcane	2303	42	85	70.2508961	100	blastp
	BQ536871_T2
957	sugarcane\|gb157.2\|	sugarcane	2304	42	83	97.4910394	92.6666667	blastp
	BU103568_T1
958	sugarcane\|gb157.2\|	sugarcane	2305	42	81	97.4910394	95.862069	blastp
	BQ536871_T1
959	sugarcane\|gb157.2\|	sugarcane	2306	42	84	97.4910394	95.862069	blastp
	BQ535332_T1
960	sunflower\|gb162\|	sunflower	2307	42	83	100	100	blastp
	CD849689_T1
961	sunflower\|gb162\|	sunflower	2308	42	83	94.9820789	95.0704225	blastp
	CD849494_T1
962	sunflower\|gb162\|	sunflower	2309	42	81	83.5125448	98.75	blastp
	CF091932_T1
963	sunflower\|gb162\|	sunflower	2310	42	81	100	100	blastp
	DY915644_T1
964	sunflower\|gb162\|	sunflower	2311	42	81	78.4946237	88.8446215	blastp
	EL462160_T1
965	sunflower\|gb162\|	sunflower	2312	42	87	95.6989247	100	blastp
	DY918039_T1
966	sunflower\|gb162\|	sunflower	2313	42	87	89.9641577	95.5223881	blastp
	DY951506_T1
967	sunflower\|gb162\|	sunflower	2314	42	84	99.6415771	98.9473684	blastp
	DY933519_T1
968	sunflower\|gb162\|	sunflower	2315	42	82	100	100	blastp
	DY917102_T1
969	sunflower\|gb162\|	sunflower	2316	42	86	58.0645161	100	blastp
	DY932916_T1
970	sunflower\|gb162\|	sunflower	2317	42	84	100	100	blastp
	CD857425_T1
971	sunflower\|gb162\|	sunflower	2318	42	85	53.4050179	97.4683544	blastp
	CD846314_T1
972	sunflower\|gb162\|	sunflower	2319	42	88	68.1003584	89.6226415	blastp
	CX944368_T1
973	sunflower\|gb162\|	sunflower	2320	42	80	100	100	blastp
	DY908592_T1
974	switchgrass\|gb165\|	switchgrass	2321	42	81	96.7741935	98.9208633	blastp
	DN141399_T1
975	switchgrass\|gb165\|	switchgrass	2322	42	86	51.9713262	99.3150685	blastp
	FE621421_T1
976	switchgrass\|gb165\|	switchgrass	2323	42	83	97.4910394	95.862069	blastp
	DN145656_T1
977	switchgrass\|gb165\|	switchgrass	2324	42	81	82.437276	100	blastp
	FE637674_T1
978	switchgrass\|gb165\|	switchgrass	2325	42	83	77.0609319	94.8497854	blastp
	DN141334_T1
979	switchgrass\|gb165\|	switchgrass	2326	42	83	85.6630824	100	blastp
	FE621578_T1
980	thellungiella\|gb157.2\|	thellungiella	2327	42	81	72.4014337	100	blastp
	DN775526_T1
981	tobacco\|gb162\|CK720599_T1	tobacco	2328	42	95	100	100	blastp
982	tobacco\|gb162\|CK720599_T2	tobacco	2329	42	94	100	100	blastp
983	tobacco\|gb162\|EB445427_T1	tobacco	2330	42	81	100	100	blastp
984	tobacco\|gb162\|EB443112_T1	tobacco	2331	42	89	100	100	blastp
985	tobacco\|gb162\|AF154641_T1	tobacco	2332	42	80	100	100	blastp
986	tobacco\|gb162\|EB425288_T1	tobacco	2333	42	94	98.2078853	100	blastp
987	tomato\|gb164\|BG123951_T2	tomato	2334	42	81	92.8315412	97.4074074	blastp
988	tomato\|gb164\|BG123951_T1	tomato	2335	42	80	98.2078853	96.8641115	blastp
989	tomato\|gb164\|BG713781_T1	tomato	2336	42	92	53.4050179	98.0519481	blastp
990	tomato\|gb164\|AW219533_T1	tomato	2337	42	81	97.1326165	98.2142857	blastp
991	triphysaria\|gb164\|	triphysaria	2338	42	88	99.2831541	99.2857143	blastp
	DR170852_T1
992	triphysaria\|gb164\|	triphysaria	2339	42	88	64.874552	100	blastp
	BM356582_T1
993	triphysaria\|gb164\|	triphysaria	2340	42	80	100	100	blastp
	BE574767_T1
994	wheat\|gb164\|BE444481_T1	wheat	2341	42	82	55.5555556	100	blastp
995	wheat\|gb164\|BE398316_T1	wheat	2342	42	81	100	100	blastp
996	wheat\|gb164\|BE404002_T1	wheat	2343	42	82	51.6129032	99.3055556	blastp
997	apple\|gb157.3\|CN892655_T1	apple	2344	43	84	100	100	blastp
998	apple\|gb157.3\|AU223658_T1	apple	2345	43	85	100	100	blastp
999	aquilegia\|gb157.3\|	aquilegia	2346	43	84	100	100	blastp
	DR914359_T1
1000	arabidopsis\|gb165\|	arabidopsis	2347	43	84	100	100	blastp
	AT2G16850_T1
1001	arabidopsis\|gb165\|	arabidopsis	2348	43	86	100	100	blastp
	AT4G35100_T1
1002	avocado\|gb164\|CK753629_T1	avocado	2349	43	85	66.4310954	92.6108374	blastp
1003	avocado\|gb164\|CK752541_T1	avocado	2350	43	84	62.1908127	100	blastp
1004	b_juncea\|gb164\|	b_juncea	2351	43	86	67.1378092	100	blastp
	EVGN01060535361904_T1
1005	b_juncea\|gb164\|	b_juncea	2352	43	86	85.1590106	100	blastp
	EVGN00138211060167_T1
1006	b_juncea\|gb164\|	b_juncea	2353	43	86	66.0777385	100	blastp
	EVGN01177009332048_T1
1007	b_juncea\|gb164\|	b_juncea	2354	43	86	90.459364	100	blastp
	EVGN00071418640425_T1
1008	b_juncea\|gb164\|	b_juncea	2355	43	92	53.0035336	100	blastp
	EVGN01391814101746_T1
1009	b_juncea\|gb164\|	b_juncea	2356	43	85	100	100	blastp
	EVGN00206114600060_T1
1010	b_oleracea\|gb161\|	b_oleracea	2357	43	85	100	100	blastp
	AF314656_T1
1011	b_oleracea\|gb161\|	b_oleracea	2358	43	86	100	100	blastp
	DY029187_T1
1012	b_oleracea\|gb161\|	b_oleracea	2359	43	87	54.770318	100	blastp
	DY014978_T1
1013	b_rapa\|gb162\|BQ791230_T1	b_rapa	2360	43	86	77.0318021	97.7375566	blastp
1014	b_rapa\|gb162\|EX033370_T1	b_rapa	2361	43	87	100	100	blastp
1015	b_rapa\|gb162\|CV432651_T1	b_rapa	2362	43	86	100	100	blastp
1016	b_rapa\|gb162\|BG543906_T1	b_rapa	2363	43	85	100	100	blastp
1017	bean\|gb164\|CB539790_T1	bean	2364	43	83	100	100	blastp
1018	beet\|gb162\|BVU60148_T1	beet	2365	43	80	100	100	blastp
1019	canola\|gb161\|DY007064_T1	canola	2366	43	86	100	100	blastp
1020	canola\|gb161\|CD815284_T1	canola	2367	43	85	100	100	blastp
1021	canola\|gb161\|CD817684_T1	canola	2368	43	85	100	100	blastp
1022	canola\|gb161\|CD821191_T1	canola	2369	43	86	100	100	blastp
1023	canola\|gb161\|CD820375_T1	canola	2370	43	87	100	100	blastp
1024	cassava\|gb164\|BM259748_T1	cassava	2371	43	84	100	100	blastp
1025	castorbean\|gb160\|	castorbean	2372	43	84	100	100	blastp
	AJ605569_T1
1026	castorbean\|gb160\|	castorbean	2373	43	84	69.9646643	95.1219512	blastp
	AJ605569_T2
1027	cichorium\|gb161\|	cichorium	2374	43	89	67.1378092	100	blastp
	EH704748_T1
1028	citrus\|gb157.2\|BQ623127_T1	citrus	2375	43	85	100	100	blastp
1029	citrus\|gb157.2\|CX664964_T1	citrus	2376	43	84	100	100	blastp
1030	clover\|gb162\|BB911260_T1	clover	2377	43	82	54.0636042	100	blastp
1031	coffea\|gb157.2\|BQ448890_T1	coffea	2378	43	86	100	100	blastp
1032	cotton\|gb164\|AI726086_T1	cotton	2379	43	84	97.8798587	88.02589	blastp
1033	cotton\|gb164\|CO098535_T1	cotton	2380	43	82	92.2261484	95.5719557	blastp
1034	cotton\|gb164\|BE051956_T1	cotton	2381	43	83	100	100	blastp
1035	cotton\|gb164\|BQ414250_T1	cotton	2382	43	85	59.0106007	100	blastp
1036	cotton\|gb164\|BF268907_T1	cotton	2383	43	87	90.1060071	100	blastp
1037	cotton\|gb164\|CO075847_T1	cotton	2384	43	84	62.5441696	100	blastp
1038	cotton\|gb164\|BQ405584_T1	cotton	2385	43	82	95.4063604	95.7142857	blastp
1039	cotton\|gb164\|AI055551_T1	cotton	2386	43	85	100	100	blastp
1040	cowpea\|gb166\|FC456786_T1	cowpea	2387	43	84	100	100	blastp
1041	dandelion\|gb161\|	dandelion	2388	43	85	100	100	blastp
	DY803814_T1
1042	ginger\|gb164\|DY347270_T1	ginger	2389	43	80	100	100	blastp
1043	ginger\|gb164\|DY347296_T1	ginger	2390	43	84	94.6996466	92.733564	blastp
1044	ginger\|gb164\|DY358056_T1	ginger	2391	43	81	100	100	blastp
1045	ginger\|gb164\|DY347277_T1	ginger	2392	43	81	100	100	blastp
1046	ginger\|gb164\|DY360032_T1	ginger	2393	43	81	81.9787986	93.902439	blastp
1047	grape\|gb160\|BM437910_T1	grape	2394	43	84	100	100	blastp
1048	iceplant\|gb164\|	iceplant	2395	43	82	100	100	blastp
	MCU26538_T1
1049	ipomoea\|gb157.2\|	ipomoea	2396	43	83	100	100	blastp
	BJ553491_T1
1050	lettuce\|gb157.2\|	lettuce	2397	43	84	100	100	blastp
	DW046509_T1
1051	lettuce\|gb157.2\|	lettuce	2398	43	84	97.8798587	100	blastp
	DW106553_T1
1052	lettuce\|gb157.2\|	lettuce	2399	43	84	100	100	blastp
	DW146059_T1
1053	lettuce\|gb157.2\|	lettuce	2400	43	83	97.1731449	100	blastp
	DW110223_T1
1054	lettuce\|gb157.2\|	lettuce	2401	43	84	100	100	blastp
	DW079482_T1
1055	lettuce\|gb157.2\|	lettuce	2402	43	83	97.5265018	92.8327645	blastp
	DW094572_T1
1056	lotus\|gb157.2\|AW163949_T1	lotus	2403	43	85	54.4169611	93.902439	blastp
1057	lotus\|gb157.2\|AI967574_T1	lotus	2404	43	82	98.5865724	97.5352113	blastp
1058	melon\|gb165\|DV632217_T1	melon	2405	43	83	100	100	blastp
1059	oil_palm\|gb166\|	oil_palm	2406	43	80	100	100	blastp
	CN600073_T1
1060	papaya\|gb165\|EX227970_T1	papaya	2407	43	85	100	100	blastp
1061	peach\|gb157.2\|AF367457_T1	peach	2408	43	85	85.1590106	99.5833333	blastp
1062	pepper\|gb157.2\|	pepper	2409	43	96	67.4911661	98.9637306	blastp
	BM066074_T1
1063	pepper\|gb157.2\|	pepper	2410	43	88	53.7102473	100	blastp
	CA518686_T1
1064	periwinkle\|gb164\|	periwinkle	2411	43	90	100	100	blastp
	AM232518_T1
1065	petunia\|gb157.2\|	petunia	2412	43	89	100	100	blastp
	AF452013_T1
1066	poplar\|gb157.2\|AI163573_T1	poplar	2413	43	84	100	100	blastp
1067	poplar\|gb157.2\|AI162424_T1	poplar	2414	43	83	100	100	blastp
1068	potato\|gb157.2\|BF053675_T1	potato	2415	43	89	83.745583	100	blastp
1069	potato\|gb157.2\|BF459952_T1	potato	2416	43	97	100	100	blastp
1070	radish\|gb164\|EV526465_T1	radish	2417	43	86	100	100	blastp
1071	radish\|gb164\|EV539317_T1	radish	2418	43	85	100	100	blastp
1072	radish\|gb164\|EV525026_T1	radish	2419	43	85	100	100	blastp
1073	rose\|gb157.2\|BQ103996_T1	rose	2420	43	82	100	100	blastp
1074	safflower\|gb162\|	safflower	2421	43	83	100	100	blastp
	EL372747_T1
1075	sesame\|gb157.2\|	sesame	2422	43	88	50.8833922	100	blastp
	BU669158_T1
1076	sesame\|gb157.2\|	sesame	2423	43	87	51.9434629	100	blastp
	BU668646_T1
1077	soybean\|gb166\|BE823128_T1	soybean	2424	43	82	100	100	blastp
1078	soybean\|gb166\|BE352716_T1	soybean	2425	43	82	100	100	blastp
1079	spruce\|gb162\|CO217407_T1	spruce	2426	43	80	93.2862191	93.5714286	blastp
1080	spurge\|gb161\|AW821924_T1	spurge	2427	43	84	100	97.2125436	blastp
1081	strawberry\|gb164\|	strawberry	2428	43	82	100	100	blastp
	CX661107_T1
1082	sunflower\|gb162\|	sunflower	2429	43	85	100	100	blastp
	CD853582_T1
1083	sunflower\|gb162\|	sunflower	2430	43	80	97.1731449	97.833935	blastp
	DY939653_T1
1084	sunflower\|gb162\|	sunflower	2431	43	81	100	100	blastp
	CD849663_T1
1085	thellungiella\|gb157.2\|	thellungiella	2432	43	86	72.0848057	90.5829596	blastp
	DN777165_T1
1086	tobacco\|gb162\|CK720587_T1	tobacco	2433	43	90	99.6466431	100	blastp
1087	tobacco\|gb162\|CK720585_T1	tobacco	2434	43	90	100	100	blastp
1088	tobacco\|gb162\|CV016422_T1	tobacco	2435	43	96	59.7173145	100	blastp
1089	tobacco\|gb162\|CK720589_T1	tobacco	2436	43	94	100	100	blastp
1090	tomato\|gb164\|BG124140_T1	tomato	2437	43	88	100	100	blastp
1091	triphysaria\|gb164\|	triphysaria	2438	43	83	100	100	blastp
	EY018490_T1
1092	triphysaria\|gb164\|	triphysaria	2439	43	84	100	100	blastp
	EY007858_T1
1093	apple\|gb157.3\|CN494428_T1	apple	2440	44	80	79.1390728	93.0232558	blastp
1094	castorbean\|gb160\|	castorbean	2441	44	82	99.0066225	97.7272727	blastp
	EG696741_T1
1095	citrus\|gb157.2\|CX074332_T1	citrus	2442	44	82	99.0066225	97.6973684	blastp
1096	citrus\|gb157.2\|CX674035_T1	citrus	2443	44	80	86.7549669	85.8085809	blastp
1097	cotton\|gb164\|DW234737_T1	cotton	2444	44	80	99.6688742	98.3333333	blastp
1098	lettuce\|gb157.2\|	lettuce	2445	44	80	95.0331126	100	blastp
	DW066284_T1
1099	petunia\|gb157.2\|	petunia	2446	44	84	65.8940397	100	blastp
	CV298254_T1
1100	potato\|gb157.2\|CV500020_T1	potato	2447	44	94	72.1854305	99.543379	blastp
1101	tobacco\|gb162\|EB426672_T1	tobacco	2448	44	89	99.0066225	97.689769	blastp
1102	brachypodium\|gb161.xeno\|	brachypodium	2449	46	92	100	100	blastp
	BE415047_T1
1103	maize\|gb164\|CF244342_T1	maize	2450	46	90	90.7630522	100	blastp
1104	maize\|gb164\|AF057183_T1	maize	2451	46	89	100	100	blastp
1105	sorghum\|gb161.xeno\|	sorghum	2452	46	88	100	100	blastp
	AF057183_T1
1106	sugarcane\|gb157.2\|	sugarcane	2453	46	89	100	100	blastp
	CA132045_T1
1107	switchgrass\|gb165\|	switchgrass	2454	46	90	100	100	blastp
	FE617713_T1
1108	wheat\|gb164\|BE430088_T1	wheat	2455	46	95	100	100	blastp
1109	apple\|gb157.3\|DT001281_T1	apple	2456	47	81	87.9518072	100	blastp
1110	brachypodium\|gb161.xeno\|	brachypodium	2457	47	95	100	100	blastp
	BE402447_T1
1111	ginger\|gb164\|DY364894_T1	ginger	2458	47	88	64.2570281	98.7654321	blastp
1112	maize\|gb164\|AI855402_T1	maize	2459	47	89	100	100	blastp
1113	maize\|gb164\|AW017703_T1	maize	2460	47	85	100	100	blastp
1114	onion\|gb162\|ACU58207_T1	onion	2461	47	83	99.5983936	99.5967742	blastp
1115	onion\|gb162\|CF437464_T1	onion	2462	47	85	95.9839357	98.75	blastp
1116	onion\|gb162\|CF435351_T1	onion	2463	47	84	100	100	blastp
1117	rice\|gb157.2\|AU031632_T1	rice	2464	47	91	100	100	blastp
1118	rice\|gb157.2\|CA756239_T1	rice	2465	47	87	97.5903614	31.640625	blastp
1119	sorghum\|gb161.xeno\|	sorghum	2466	47	89	100	100	blastp
	AI855402_T1
1120	sugarcane\|gb157.2\|	sugarcane	2467	47	90	99.5983936	99.5967742	blastp
	CA132045_T2
1121	switchgrass\|gb165\|	switchgrass	2468	47	90	100	100	blastp
	FE624217_T1
1122	wheat\|gb164\|BE404100_T1	wheat	2469	47	97	100	100	blastp
1123	fescue\|gb161\|DT700572_T1	fescue	2470	48	94	100	100	blastp
1124	ginger\|gb164\|DY361836_T1	ginger	2471	48	80	95.1612903	96.7346939	blastp
1125	rice\|gb157.2\|U37952_T1	rice	2472	48	90	100	100	blastp
1126	rice\|gb157.2\|U37952_T3	rice	2473	48	89	51.6129032	89.5104895	blastp
1127	rye\|gb164\|BE495605_T1	rye	2474	48	98	80.6451613	100	blastp
1128	sorghum\|gb161.xeno\|	sorghum	2475	48	90	100	100	blastp
	AI724211_T1
1129	sugarcane\|gb157.2\|	sugarcane	2476	48	82	92.3387097	100	blastp
	CA110414_T1
1130	sugarcane\|gb157.2\|	sugarcane	2477	48	83	90.3225806	96.5517241	blastp
	CA065356_T1
1131	sugarcane\|gb157.2\|	sugarcane	2478	48	83	71.3709677	95.6756757	blastp
	CA143208_T1
1132	sugarcane\|gb157.2\|	sugarcane	2479	48	91	100	100	blastp
	CA101765_T1
1133	sugarcane\|gb157.2\|	sugarcane	2480	48	90	83.4677419	95.8333333	blastp
	CA133231_T1
1134	switchgrass\|gb165\|	switchgrass	2481	48	91	100	100	blastp
	FE605472_T1
1135	wheat\|gb164\|BE489764_T1	wheat	2482	48	95	100	100	blastp
1136	wheat\|gb164\|TAU86763_T1	wheat	2483	48	95	100	100	blastp
1137	wheat\|gb164\|BE415001_T1	wheat	2484	48	95	100	100	blastp
1138	b_juncea\|gb164\|	b_juncea	2485	50	80	85.0931677	100	blastp
	EVGN00504508791211_T1
1139	b_juncea\|gb164\|	b_juncea	2486	50	84	50.931677	91.954023	blastp
	EVGN01684214261870_T1
1140	banana\|gb160\|DN239388_T1	banana	2487	50	84	80.1242236	91.9708029	blastp
1141	barley\|gb157.3\|BF253694_T1	barley	2488	50	91	58.3850932	98.9690722	blastp
1142	barley\|gb157.3\|BE412959_T3	barley	2489	50	95	100	62.6923077	blastp
1143	bean\|gb164\|FD793482_T1	bean	2490	50	82	100	91.9075145	blastp
1144	canola\|gb161\|CX193398_T3	canola	2491	50	83	99.378882	66.9527897	blastp
1145	canola\|gb161\|EV123336_T1	canola	2492	50	81	52.7950311	91.2087912	blastp
1146	centaurea\|gb161\|	centaurea	2493	50	81	98.136646	62.248996	blastp
	EL931277_T1
1147	cichorium\|gb161\|	cichorium	2494	50	83	64.5962733	95.3271028	blastp
	DT213939_T1
1148	fescue\|gb161\|DT703843_T1	fescue	2495	50	99	100	94.1520468	blastp
1149	lotus\|gb157.2\|BI418499_T1	lotus	2496	50	86	98.136646	88.700565	blastp
1150	onion\|gb162\|BQ579939_T1	onion	2497	50	86	100	57.1942446	blastp
1151	peach\|gb157.2\|BU040795_T1	peach	2498	50	82	98.136646	56.2043796	blastp
1152	peach\|gb157.2\|DW351857_T1	peach	2499	50	83	98.136646	91.8128655	blastp
1153	rye\|gb164\|BF429463_T1	rye	2500	50	88	93.1677019	100	blastp
1154	soybean\|gb166\|BE352747_T4	soybean	2501	50	81	98.136646	70.7207207	blastp
1155	sugarcane\|gb157.2\|	sugarcane	2502	50	81	98.136646	56.6787004	blastp
	CA194640_T1
1156	sugarcane\|gb157.2\|	sugarcane	2503	50	84	96.2732919	87.0056497	blastp
	CA103332_T1
1157	sugarcane\|gb157.2\|	sugarcane	2504	50	82	93.7888199	74.2574257	blastp
	CA167616_T1
1158	sugarcane\|gb157.2\|	sugarcane	2505	50	90	100	68.5344828	blastp
	CA103740_T1
1159	sugarcane\|gb157.2\|	sugarcane	2506	50	82	97.515528	77.4834437	tblastn
	CA184547_T1
1160	sunflower\|gb162\|	sunflower	2507	50	87	98.136646	87.5	blastp
	CF089373_T1
1161	wheat\|gb164\|CA484201_T1	wheat	2508	50	89	96.8944099	65.9574468	blastp
1162	apricot\|gb157.2\|	apricot	2509	51	86	100	65.9574468	blastp
	CV049856_T1
1163	arabidopsis\|gb165\|	arabidopsis	2510	51	89	100	32.6315789	blastp
	AT2G37180_T1
1164	arabidopsis\|gb165\|	arabidopsis	2511	51	92	100	32.1799308	blastp
	\|AT2G39010_T1
1165	b_juncea\|gb164\|	b_juncea	2512	51	87	100	86.9158879	blastp
	EVGN00130608921231_T1
1166	b_juncea\|gb164\|	b_juncea	2513	51	86	90.3225806	86.5979381	blastp
	EVGN00605203140273_T1
1167	b_juncea\|gb164\|	b_juncea	2514	51	86	56.9892473	91.3793103	blastp
	EVGN21262514941904_T1
1168	b_juncea\|gb164\|	b_juncea	2515	51	86	100	85.3211009	blastp
	EVGN00733314152324_T1
1169	b_juncea\|gb164\|	b_juncea	2516	51	86	89.2473118	49.4047619	blastp
	EVGN00041211340240_T1
1170	b_juncea\|gb164\|	b_juncea	2517	51	91	100	80.1724138	blastp
	DT317704_T1
1171	b_juncea\|gb164\|	b_juncea	2518	51	85	89.2473118	79.8076923	blastp
	EVGN00208014701957_T1
1172	b_juncea\|gb164\|	b_juncea	2519	51	90	100	36.328125	blastp
	EVGN00550314491066_T1
1173	b_juncea\|gb164\|	b_juncea	2520	51	90	90.3225806	95.4545455	blastp
	EVGN01267508262672_T1
1174	b_juncea\|gb164\|	b_juncea	2521	51	89	100	51.3812155	blastp
	EVGN01803715320789_T1
1175	b_juncea\|gb164\|	b_juncea	2522	51	91	100	34.4019729	tblastn
	EVGN00397011681539_T1
1176	b_rapa\|gb162\|DN965016_T1	b_rapa	2523	51	88	100	69.4029851	blastp
1177	b_rapa\|gb162\|EX025548_T1	b_rapa	2524	51	87	100	32.5174825	blastp
1178	b_rapa\|gb162\|BG543764_T1	b_rapa	2525	51	92	100	32.2916667	blastp
1179	banana\|gb160\|DN238638_T1	banana	2526	51	89	100	27.8721279	tblastn
1180	banana\|gb160\|DN238638_T2	banana	2527	51	89	100	30.5921053	tblastn
1181	barley\|gb157.3\|BE421292_T1	barley	2528	51	87	100	32.7464789	blastp
1182	barley\|gb157.3\|BJ446923_T1	barley	2529	51	83	100	64.5833333	blastp
1183	beet\|gb162\|BQ488455_T1	beet	2530	51	81	100	26.686217	blastp
1184	beet\|gb162\|BVU60147_T1	beet	2531	51	84	100	32.2916667	blastp
1185	canola\|gb161\|CX189721_T1	canola	2532	51	87	100	32.5174825	blastp
1186	canola\|gb161\|CB686155_T1	canola	2533	51	92	100	32.2916667	blastp
1187	canola\|gb161\|DY007249_T1	canola	2534	51	87	100	32.5174825	blastp
1188	canola\|gb161\|ES986486_T1	canola	2535	51	86	96.7741935	87.3786408	blastp
1189	canola\|gb161\|EV203446_T1	canola	2536	51	82	100	38.0108992	tblastn
1190	cassava\|gb164\|DN740353_T1	cassava	2537	51	93	100	62.8378378	blastp
1191	cassava\|gb164\|DV449516_T1	cassava	2538	51	94	100	68.8888889	blastp
1192	cassava\|gb164\|BM259717_T1	cassava	2539	51	81	100	32.8621908	blastp
1193	castorbean\|gb160\|	castorbean	2540	51	81	100	46.5	blastp
	EE257493_T2
1194	castorbean\|gb160\|	castorbean	2541	51	81	100	34.4444444	blastp
	EE257493_T1
1195	cichorium\|gb161\|	cichorium	2542	51	94	100	80.8695652	blastp
	EH706421_T1
1196	cichorium\|gb161\|	cichorium	2543	51	90	100	32.5554259	tblastn
	EH708948_T1
1197	cichorium\|gb161\|	cichorium	2544	51	90	100	24.668435	tblastn
	EH692078_T1
1198	citrus\|gb157.2\|CB291468_T1	citrus	2545	51	82	100	86.1111111	blastp
1199	citrus\|gb157.2\|BQ624699_T1	citrus	2546	51	90	100	27.56917	tblastn
1200	clover\|gb162\|BB903718_T1	clover	2547	51	91	100	32.6315789	blastp
1201	clover\|gb162\|BB930902_T1	clover	2548	51	88	100	32.4041812	blastp
1202	clover\|gb162\|BB911526_T1	clover	2549	51	91	92.4731183	80.3738318	blastp
1203	clover\|gb162\|BB913405_T1	clover	2550	51	90	96.7741935	81.0810811	blastp
1204	cotton\|gb164\|ES804497_T1	cotton	2551	51	86	100	58.490566	blastp
1205	cotton\|gb164\|EX172153_T1	cotton	2552	51	89	90.3225806	41.5156507	tblastn
1206	cowpea\|gb166\|FF384339_T1	cowpea	2553	51	89	90.3225806	80	blastp
1207	cowpea\|gb166\|FF396241_T1	cowpea	2554	51	82	97.8494624	88.3495146	blastp
1208	cowpea\|gb166\|ES884224_T1	cowpea	2555	51	88	100	32.4041812	blastp
1209	cowpea\|gb166\|FG857474_T1	cowpea	2556	51	89	100	68.8888889	blastp
1210	cryptomeria\|gb166\|	cryptomeria	2557	51	81	100	78.8135593	blastp
	BY902595_T1
1211	cryptomeria\|gb166\|	cryptomeria	2558	51	83	100	31.7406143	blastp
	BJ937695_T1
1212	cryptomeria\|gb166\|	cryptomeria	2559	51	82	88.172043	86.3157895	blastp
	BW993227_T1
1213	dandelion\|gb161\|	dandelion	2560	51	87	91.3978495	75.2212389	blastp
	DY802675_T1
1214	fescue\|gb161\|DT674412_T1	fescue	2561	51	82	84.9462366	85.8695652	blastp
1215	fescue\|gb161\|DT695652_T1	fescue	2562	51	84	100	76.8595041	blastp
1216	fescue\|gb161\|DT702501_T1	fescue	2563	51	83	95.6989247	96.7391304	blastp
1217	fescue\|gb161\|DT688112_T1	fescue	2564	51	83	100	85.3211009	blastp
1218	fescue\|gb161\|DT688728_T1	fescue	2565	51	92	100	81.5789474	blastp
1219	ginger\|gb164\|DY358169_T1	ginger	2566	51	87	100	32.9787234	blastp
1220	ginger\|gb164\|DY366672_T1	ginger	2567	51	82	93.5483871	87	blastp
1221	ginger\|gb164\|DY360033_T1	ginger	2568	51	91	100	27.7888446	tblastn
1222	grape\|gb160\|AF188843_T5	grape	2569	51	82	100	32.5174825	blastp
1223	ipomoea\|gb157.2\|	ipomoea	2570	51	88	100	32.4041812	blastp
	BJ556470_T1
1224	lettuce\|gb157.2\|	lettuce	2571	51	90	100	32.6315789	blastp
	DW158018_T1
1225	lettuce\|gb157.2\|	lettuce	2572	51	90	100	34.7014925	blastp
	DW091407_T1
1226	lettuce\|gb157.2\|	lettuce	2573	51	89	89.2473118	32.6771654	blastp
	DW060777_T1
1227	lotus\|gb157.2\|AV775277_T1	lotus	2574	51	83	100	88.5714286	blastp
1228	lotus\|gb157.2\|AI967387_T1	lotus	2575	51	84	100	32.4041812	blastp
1229	lotus\|gb157.2\|AV774377_T1	lotus	2576	51	81	100	88.5714286	blastp
1230	lotus\|gb157.2\|BP059122_T1	lotus	2577	51	80	73.1182796	85	blastp
1231	lotus\|gb157.2\|BI419853_T1	lotus	2578	51	81	94.6236559	88	blastp
1232	lotus\|gb157.2\|BP049219_T1	lotus	2579	51	81	100	76.2295082	blastp
1233	lotus\|gb157.2\|AV775053_T1	lotus	2580	51	82	84.9462366	86.8131868	blastp
1234	lotus\|gb157.2\|AV775249_T1	lotus	2581	51	85	95.6989247	80.9090909	blastp
1235	maize\|gb164\|AF326496_T1	maize	2582	51	88	100	32.4041812	blastp
1236	maize\|gb164\|AY107589_T1	maize	2583	51	84	100	32.8621908	blastp
1237	medicago\|gb157.2\|	medicago	2584	51	87	100	32.4041812	blastp
	AI974409_T1
1238	medicago\|gb157.2\|	medicago	2585	51	89	100	32.6315789	blastp
	AI974231_T1
1239	melon\|gb165\|EB715587_T1	melon	2586	51	86	100	23.4848485	tblastn
1240	oat\|gb164\|CN816056_T1	oat	2587	51	82	100	84.5454545	blastp
1241	oil_palm\|gb166\|	oil_palm	2588	51	89	100	32.9787234	blastp
	CN601069_T1
1242	oil_palm\|gb166\|	oil_palm	2589	51	84	100	32.9787234	blastp
	EL686181_T1
1243	peanut\|gb161\|CD037823_T1	peanut	2590	51	82	100	31.8493151	blastp
1244	peanut\|gb161\|CD038014_T1	peanut	2591	51	88	100	32.1799308	blastp
1245	periwinkle\|gb164\|	periwinkle	2592	51	87	100	65.9574468	blastp
	AM232518_T2
1246	physcomitrella\|gb157\|	physcomitrella	2593	51	89	100	33.3333333	blastp
	BI436955_T1
1247	physcomitrella\|gb157\|	physcomitrella	2594	51	82	100	32.5174825	blastp
	BJ198543_T1
1248	physcomitrella\|gb157\|	physcomitrella	2595	51	90	100	33.2142857	blastp
	AW476973_T3
1249	physcomitrella\|gb157\|	physcomitrella	2596	51	82	100	32.5259516	blastp
	BJ962015_T1
1250	pine\|gb157.2\|AW870138_T1	pine	2597	51	82	100	34.7014925	blastp
1251	pine\|gb157.2\|AI813147_T1	pine	2598	51	87	100	33.0960854	blastp
1252	pine\|gb157.2\|AA739836_T1	pine	2599	51	87	100	33.0960854	blastp
1253	pine\|gb157.2\|AL750425_T1	pine	2600	51	88	100	33.0960854	blastp
1254	pine\|gb157.2\|BG038984_T1	pine	2601	51	80	100	32.8621908	blastp
1255	pine\|gb157.2\|AW225939_T1	pine	2602	51	80	100	34.1911765	blastp
1256	pine\|gb157.2\|BQ696500_T1	pine	2603	51	81	100	32.8621908	blastp
1257	pine\|gb157.2\|AL751198_T1	pine	2604	51	87	100	33.3333333	blastp
1258	pine\|gb157.2\|BQ695693_T1	pine	2605	51	82	100	32.8621908	blastp
1259	pine\|gb157.2\|BF517326_T1	pine	2606	51	80	100	60.3896104	blastp
1260	pine\|gb157.2\|AA739625_T1	pine	2607	51	81	100	34.7014925	blastp
1261	pine\|gb157.2\|BG317873_T1	pine	2608	51	82	100	32.8621908	blastp
1262	pine\|gb157.2\|AI813147_T2	pine	2609	51	87	100	32.9787234	blastp
1263	pine\|gb157.2\|CF388120_T1	pine	2610	51	87	100	33.3333333	blastp
1264	pine\|gb157.2\|BE662590_T1	pine	2611	51	81	100	35.7692308	blastp
1265	pine\|gb157.2\|BG318657_T1	pine	2612	51	82	100	32.8621908	blastp
1266	pine\|gb157.2\|AA557104_T1	pine	2613	51	88	100	33.3333333	blastp
1267	pine\|gb157.2\|AW870138_T2	pine	2614	51	82	100	32.8621908	blastp
1268	pine\|gb157.2\|AW289749_T1	pine	2615	51	82	100	32.8621908	blastp
1269	pine\|gb157.2\|H75016_T1	pine	2616	51	88	100	32.7464789	blastp
1270	pine\|gb157.2\|AA740005_T1	pine	2617	51	80	100	35.0943396	blastp
1271	pine\|gb157.2\|CA305579_T1	pine	2618	51	91	100	75.6097561	blastp
1272	pine\|gb157.2\|BE187350_T1	pine	2619	51	83	100	32.8621908	blastp
1273	pine\|gb157.2\|CF473539_T1	pine	2620	51	84	100	32.9787234	blastp
1274	pine\|gb157.2\|AW290370_T1	pine	2621	51	83	100	32.7464789	blastp
1275	pine\|gb157.2\|AW290691_T1	pine	2622	51	82	75.2688172	88.6075949	blastp
1276	pine\|gb157.2\|BG318695_T1	pine	2623	51	82	100	34.7014925	blastp
1277	pineapple\|gb157.2\|	pineapple	2624	51	91	98.9247312	78.6324786	blastp
	DT339628_T1
1278	poplar\|gb157.2\|AI162483_T2	poplar	2625	51	86	100	24.3455497	tblastn
1279	poplar\|gb157.2\|BU817536_T4	poplar	2626	51	87	100	22.6094003	tblastn
1280	potato\|gb157.2\|BQ515617_T1	potato	2627	51	80	98.9247312	36.6533865	blastp
1281	potato\|gb157.2\|BE920139_T2	potato	2628	51	90	100	41.5178571	blastp
1282	radish\|gb164\|EV526963_T1	radish	2629	51	87	100	32.5174825	blastp
1283	radish\|gb164\|EV567230_T2	radish	2630	51	84	100	46.039604	blastp
1284	radish\|gb164\|EV551004_T1	radish	2631	51	92	100	32.2916667	blastp
1285	radish\|gb164\|EX756464_T1	radish	2632	51	86	100	48.1865285	blastp
1286	radish\|gb164\|EY910551_T1	radish	2633	51	91	100	31.9587629	blastp
1287	radish\|gb164\|EW730466_T1	radish	2634	51	84	98.9247312	87.6190476	blastp
1288	radish\|gb164\|AF051128_T1	radish	2635	51	81	87.0967742	48.6	tblastn
1289	radish\|gb164\|EX756028_T1	radish	No	51	86	100	40.0286944	tblastn
			predicted
			Protein
1290	rice\|gb157.2\|BE229418_T1	rice	2636	51	90	100	32.9787234	blastp
1291	rice\|gb157.2\|BE229418_T3	rice	2637	51	90	100	38.5892116	blastp
1292	rice\|gb157.2\|AK107700_T1	rice	2638	51	91	100	68.8888889	blastp
1293	rice\|gb157.2\|BE229418_T4	rice	2639	51	90	100	54.0697674	blastp
1294	rice\|gb157.2\|NM001066078_T1	rice	2640	51	92	100	41.7040359	blastp
1295	rice\|gb157.2\|AW155505_T1	rice	2641	51	83	100	32.0689655	blastp
1296	rice\|gb157.2\|AW155505_T2	rice	2642	51	83	98.9247312	35.7976654	blastp
1297	rice\|gb157.2\|AK106746_T1	rice	2643	51	83	100	22.6461039	tblastn
1298	sorghum\|gb161.xeno\|	sorghum	2644	51	86	100	33.3333333	blastp
	AY107589_T1
1299	sorghum\|gb161.xeno\|	sorghum	2645	51	92	100	32.5174825	blastp
	AI947598_T1
1300	sorghum\|gb161.xeno\|	sorghum	2646	51	80	100	31.4189189	blastp
	AI855413_T1
1301	sorghum\|gb161.xeno\|	sorghum	2647	51	81	100	31.1418685	blastp
	AW565915_T1
1302	sorghum\|gb161.xeno\|	sorghum	2648	51	90	100	79.6610169	blastp
	CF481617_T1
1303	soybean\|gb166\|BU549322_T1	soybean	2649	51	89	100	32.5174825	blastp
1304	soybean\|gb166\|BE352729_T1	soybean	2650	51	88	100	32.4041812	blastp
1305	soybean\|gb166\|BI974981_T1	soybean	2651	51	91	100	71.5384615	blastp
1306	soybean\|gb166\|BE658685_T1	soybean	2652	51	89	100	32.6315789	blastp
1307	soybean\|gb166\|BE352747_T3	soybean	2653	51	81	100	20.5904059	tblastn
1308	spikemoss\|gb165\|	spikemoss	2654	51	88	100	32.0689655	blastp
	DN838269_T1
1309	spikemoss\|gb165\|	spikemoss	2655	51	88	100	32.0689655	blastp
	FE434019_T1
1310	spruce\|gb162\|CO225164_T1	spruce	2656	51	89	100	32.8621908	blastp
1311	spruce\|gb162\|CO258147_T1	spruce	2657	51	81	100	33.8181818	blastp
1312	spruce\|gb162\|CO230791_T1	spruce	2658	51	88	100	46.039604	blastp
1313	spruce\|gb162\|DR546674_T1	spruce	2659	51	82	100	32.5	blastp
1314	spruce\|gb162\|DR560237_T1	spruce	2660	51	81	100	34.1911765	blastp
1315	spurge\|gb161\|DV116550_T1	spurge	2661	51	83	100	86.1111111	blastp
1316	sugarcane\|gb157.2\|	sugarcane	2662	51	87	88.172043	81.1881188	blastp
	CA088464_T1
1317	sugarcane\|gb157.2\|	sugarcane	2663	51	92	100	62.4161074	blastp
	CA086583_T1
1318	sugarcane\|gb157.2\|	sugarcane	2664	51	86	62.3655914	100	blastp
	CA234165_T1
1319	sugarcane\|gb157.2\|	sugarcane	2665	51	89	100	32.5174825	blastp
	CA107998_T1
1320	sugarcane\|gb157.2\|	sugarcane	2666	51	95	100	34.1911765	tblastn
	CA204327_T1
1321	sugarcane\|gb157.2\|	sugarcane	2667	51	92	86.0215054	18.6480186	tblastn
	CA067786_T1
1322	sunflower\|gb162\|	sunflower	2668	51	81	100	31.9587629	blastp
	DY944685_T1
1323	sunflower\|gb162\|	sunflower	2669	51	87	52.688172	100	blastp
	BQ968590_T1
1324	sunflower\|gb162\|	sunflower	2670	51	89	98.9247312	41.3173653	tblastn
	CD848850_T1
1325	tobacco\|gb162\|EB445876_T1	tobacco	2671	51	90	100	32.4041812	blastp
1326	tobacco\|gb162\|EB445188_T1	tobacco	2672	51	88	100	32.4041812	blastp
1327	tobacco\|gb162\|CK720586_T1	tobacco	2673	51	81	100	34.7014925	blastp
1328	tomato\|gb164\|CO750818_T1	tomato	2674	51	92	88.172043	82.8282828	blastp
1329	tomato\|gb164\|AI772191_T1	tomato	2675	51	81	98.9247312	33.6996337	blastp
1330	tomato\|gb164\|AW219533_T2	tomato	2676	51	89	100	39.2405063	tblastn
1331	triphysaria\|gb164\|	triphysaria	2677	51	88	95.6989247	80.9090909	blastp
	DR173305_T1
1332	triphysaria\|gb164\|	triphysaria	2678	51	90	100	75.6097561	blastp
	EX984185_T1
1333	triphysaria\|gb164\|	triphysaria	2679	51	91	100	67.8832117	blastp
	DR174019_T1
1334	wheat\|gb164\|CA609068_T1	wheat	2680	51	94	100	67.3913043	blastp
1335	wheat\|gb164\|BE430411_T1	wheat	2681	51	86	100	32.4041812	blastp
1336	wheat\|gb164\|CK208980_T1	wheat	2682	51	80	100	30	blastp
1337	wheat\|gb164\|CA620158_T1	wheat	2683	51	96	95.6989247	87.254902	blastp
1338	wheat\|gb164\|BE405395_T1	wheat	2684	51	88	100	32.7464789	blastp
1339	wheat\|gb164\|CA647310_T1	wheat	2685	51	80	77.4193548	93.5064935	blastp
1340	wheat\|gb164\|BE492099_T1	wheat	2686	51	87	100	32.7464789	blastp
1341	wheat\|gb164\|BE402029_T1	wheat	2687	51	87	100	32.7464789	blastp
1342	wheat\|gb164\|CA618130_T1	wheat	2688	51	86	88.172043	73.2142857	blastp
1343	wheat\|gb164\|CJ605707_T1	wheat	2689	51	82	100	69.4029851	blastp
1344	wheat\|gb164\|CA614209_T1	wheat	2690	51	98	59.1397849	98.2142857	blastp
1345	wheat\|gb164\|CA701714_T1	wheat	2691	51	82	100	77.5	blastp
1346	wheat\|gb164\|BE403921_T1	wheat	2692	51	93	96.7741935	82.5688073	blastp
1347	wheat\|gb164\|BE217049_T1	wheat	2693	51	83	100	31.9587629	blastp
1348	wheat\|gb164\|CA602649_T1	wheat	2694	51	84	100	30.0322928	tblastn
1349	wheat\|gb164\|CA486220_T1	wheat	2695	51	82	96.7741935	33.3759591	tblastn
1350	b_juncea\|gb164\|	b_juncea	2696	52	85	64.3835616	99.2957746	blastp
	EVGN00044413933329_T1
1351	b_juncea\|gb164\|	b_juncea	2697	52	90	53.4246575	99.1525424	blastp
	EVGN00137910990746_T1
1352	barley\|gb157.3\|BE413268_T1	barley	2698	52	81	96.803653	73.4482759	blastp
1353	barley\|gb157.3\|AJ433979_T1	barley	2699	52	81	96.803653	73.4482759	blastp
1354	barley\|gb157.3\|BE412979_T1	barley	2700	52	82	96.803653	74.1258741	blastp
1355	brachypodium\|gb161.xeno\|	brachypodium	2701	52	83	96.803653	73.6111111	blastp
	AF139815_T1
1356	citrus\|gb157.2\|CX052950_T1	citrus	2702	52	84	50.2283105	100	blastp
1357	clover\|gb162\|BB913131_T1	clover	2703	52	93	52.5114155	99.137931	blastp
1358	clover\|gb162\|BB918704_T1	clover	2704	52	82	63.9269406	99.2957746	blastp
1359	cotton\|gb164\|CO103246_T1	cotton	2705	52	91	56.6210046	93.9393939	blastp
1360	cowpea\|gb166\|FG883860_T1	cowpea	2706	52	83	51.1415525	99.1150442	blastp
1361	fescue\|gb161\|DT702489_T1	fescue	2707	52	96	57.0776256	93.2835821	blastp
1362	fescue\|gb161\|DT702846_T1	fescue	2708	52	96	57.0776256	96.8992248	blastp
1363	ipomoea\|gb157.2\|	ipomoea	2709	52	81	87.2146119	74.7035573	blastp
	BU691146_T1
1364	maize\|gb164\|AI939909_T1	maize	2710	52	80	96.803653	73.6111111	blastp
1365	maize\|gb164\|AF130975_T1	maize	2711	52	83	96.803653	74.3859649	blastp
1366	maize\|gb164\|AI947831_T1	maize	2712	52	83	96.803653	73.6111111	blastp
1367	oil_palm\|gb166\|	oil_palm	2713	52	80	96.803653	75.177305	blastp
	EL692065_T1
1368	physcomitrella\|gb157\|	physcomitrella	2714	52	80	93.6073059	73.4767025	blastp
	AW476973_T1
1369	physcomitrella\|gb157\|	physcomitrella	2715	52	80	96.347032	73.0103806	blastp
	BI894596_T1
1370	physcomitrella\|gb157\|	physcomitrella	2716	52	80	93.6073059	73.4767025	blastp
	AW476973_T2
1371	pine\|gb157.2\|CF387570_T1	pine	2717	52	88	51.1415525	99.1150442	blastp
1372	radish\|gb164\|EY904434_T2	radish	2718	52	88	54.3378995	99.1666667	blastp
1373	radish\|gb164\|FD960377_T1	radish	2719	52	84	63.0136986	99.2805755	blastp
1374	rice\|gb157.2\|AU093957_T1	rice	2720	52	81	96.803653	74.1258741	blastp
1375	rice\|gb157.2\|BE039992_T2	rice	2721	52	92	57.0776256	97.65625	blastp
1376	rice\|gb157.2\|BE530955_T1	rice	2722	52	80	96.803653	73.1034483	blastp
1377	rye\|gb164\|BE586469_T1	rye	2723	52	82	61.1872146	97.810219	blastp
1378	sorghum\|gb161.xeno\|	sorghum	2724	52	83	96.803653	72.6027397	blastp
	AW922622_T1
1379	sorghum\|gb161.xeno\|	sorghum	2725	52	80	85.3881279	74.8	blastp
	BE344582_T1
1380	sorghum\|gb161.xeno\|	sorghum	2726	52	82	96.803653	73.3564014	blastp
	AI855280_T1
1381	spikemoss\|gb165\|	spikemoss	2727	52	83	91.7808219	71.5302491	blastp
	DN838148_T1
1382	spikemoss\|gb165\|	spikemoss	2728	52	83	91.7808219	71.5302491	blastp
	DN838057_T1
1383	sugarcane\|gb157.2\|	sugarcane	2729	52	82	90.4109589	71.8978102	blastp
	CA139573_T1
1384	sugarcane\|gb157.2\|	sugarcane	2730	52	90	57.0776256	99.2063492	blastp
	CA145403_T1
1385	sunflower\|gb162\|	sunflower	2731	52	80	57.0776256	93.2835821	blastp
	CF083179_T1
1386	sunflower\|gb162\|	sunflower	2732	52	86	56.6210046	89.2086331	blastp
	BU035823_T1
1387	switchgrass\|gb165\|	switchgrass	2733	52	83	61.6438356	99.2647059	blastp
	DN140790_T1
1388	switchgrass\|gb165\|	switchgrass	2734	52	81	66.2100457	96.6666667	blastp
	FE631354_T1
1389	tobacco\|gb162\|DV159802_T1	tobacco	2735	52	81	87.6712329	48.7722269	tblastn
1390	wheat\|gb164\|AF139815_T1	wheat	2736	52	82	96.803653	74.1258741	blastp
1391	wheat\|gb164\|BE406301_T1	wheat	2737	52	81	96.803653	73.4482759	blastp
1392	wheat\|gb164\|BE404904_T1	wheat	2738	52	81	96.803653	73.4482759	blastp
1393	wheat\|gb164\|BE400219_T1	wheat	2739	52	81	96.803653	73.4482759	blastp
1394	wheat\|gb164\|CA619093_T1	wheat	2740	52	81	54.7945205	90.1515152	blastp
1395	wheat\|gb164\|BE605056_T1	wheat	2741	52	88	56.6210046	93.2330827	blastp
1396	wheat\|gb164\|BQ245211_T1	wheat	2742	52	82	96.803653	74.1258741	blastp
1397	wheat\|gb164\|BE497487_T1	wheat	2743	52	81	96.803653	73.4482759	blastp
1398	wheat\|gb164\|BQ295206_T1	wheat	2744	52	89	64.8401826	100	blastp
1399	wheat\|gb164\|BE499954_T1	wheat	2745	52	80	98.630137	74.8275862	blastp
1400	wheat\|gb164\|BE405794_T1	wheat	2746	52	81	96.803653	73.4482759	blastp
6	tomato\|gb164\|AW934056_T1	tomato	32	31	82	100	100	blastp
21	barley\|gb157.3\|AL501410_T1	barley	47	46	87	98.3935743	98.79518072	blastp
2844	antirrhinum\|gb166\|	antirrhinum	3052	25	88	100	100	blastp
	AJ559427_T1
2845	antirrhinum\|gb166\|	antirrhinum	3053	25	85	100	100	blastp
	AJ791214_T1
2846	bruguiera\|gb166\|	bruguiera	3054	25	82	61.29032258	100	blastp
	BP939664_T1
2847	centaurea\|gb166\|	centaurea	3055	25	84	98.79032258	100	blastp
	EL931601_T1
2848	eucalyptus\|gb166\|	eucalyptus	3056	25	85	98.79032258	98.79032258	blastp
	CD668425_T1
2849	kiwi\|gb166\|FG409998_T1	kiwi	3057	25	85	100	100	blastp
2850	kiwi\|gb166\|FG453166_T1	kiwi	3058	25	86	100	100	blastp
2851	kiwi\|gb166\|FG401585_T1	kiwi	3059	25	87	99.59677419	100	blastp
2852	kiwi\|gb166\|FG419790_T1	kiwi	3060	25	85	100	100	blastp
2853	liriodendron\|gb166\|	liriodendron	3061	25	82	99.19354839	98.79518072	blastp
	FD495170_T1
2854	poppy\|gb166\|FG607362_T1	poppy	3062	25	82	82.66129032	99.51456311	blastp
2855	soybean\|gb167\|AA660186_T1	soybean	3063	25	83	100	100	blastp
2856	soybean\|gb167\|CA990807_T1	soybean	3064	25	82	95.56451613	100	blastp
2857	walnuts\|gb166\|CV196664_T1	walnuts	3065	25	83	100	100	blastp
2858	antirrhinum\|gb166\|	antirrhinum	3066	26	90	100	100	blastp
	X70417_T1
2859	banana\|gb167\|FF560721_T1	banana	3067	26	86	77.6	99.48717949	blastp
2860	centaurea\|gb166\|	centaurea	3068	26	86	100	100	blastp
	EL932548_T1
2861	antirrhinum\|gb166\|	antirrhinum	3069	27	86	90.90909091	100	blastp
	AJ789802_T1
2862	bruguiera\|gb166\|	bruguiera	3070	27	81	69.96047431	100	blastp
	BP938735_T1
2863	eucalyptus\|gb166\|	eucalyptus	3071	27	82	90.90909091	100	blastp
	ES589574_T1
2864	kiwi\|gb166\|FG427735_T1	kiwi	3072	27	86	50.19762846	99.21875	blastp
2865	kiwi\|gb166\|FG406415_T1	kiwi	3073	27	81	54.15019763	100	blastp
2866	kiwi\|gb166\|FG406885_T1	kiwi	3074	27	84	99.20948617	99.6031746	blastp
2867	liriodendron\|gb166\|	liriodendron	3075	27	81	99.20948617	99.6031746	blastp
	CK744430_T1
2868	poppy\|gb166\|FG608493_T1	poppy	3076	27	84	83.00395257	99.52606635	blastp
2869	soybean\|gb167\|EV269611_T1	soybean	3077	27	86	60.07905138	96.20253165	blastp
2870	amborella\|gb166\|	amborella	3078	28	82	100	100	blastp
	CD482678_T1
2871	cenchrus\|gb166\|	cenchrus	3079	28	81	100	100	blastp
	BM084541_T1
2872	leymus\|gb166\|EG376267_T1	leymus	3080	28	80	100	100	blastp
2873	leymus\|gb166\|EG386149_T1	leymus	3081	28	80	100	100	blastp
2874	walnuts\|gb166\|EL895384_T1	walnuts	3082	28	83	82	94.49541284	blastp
2875	amborella\|gb166\|	amborella	3083	30	88	98.26388889	98.95470383	blastp
	CD481950_T1
2876	antirrhinum\|gb166\|	antirrhinum	3084	30	86	93.40277778	100	blastp
	AJ796874_T1
2877	antirrhinum\|gb166\|	antirrhinum	3085	30	84	73.26388889	99.52606635	blastp
	AJ798039_T1
2878	antirrhinum\|gb166\|	antirrhinum	3086	30	88	85.06944444	100	blastp
	AJ792331_T1
2879	antirrhinum\|gb166\|	antirrhinum	3087	30	86	98.26388889	99.3006993	blastp
	AJ558770_T1
2880	banana\|gb167\|FF558844_T1	banana	3088	30	88	98.26388889	98.95104895	blastp
2881	bean\|gb167\|CA898412_T1	bean	3089	30	81	98.26388889	99.30313589	blastp
2882	bruguiera\|gb166\|	bruguiera	3090	30	86	52.77777778	95	blastp
	BP941115_T1
2883	bruguiera\|gb166\|	bruguiera	3091	30	85	97.91666667	99.30313589	blastp
	BP939033_T1
2884	cenchrus\|gb166\|	cenchrus	3092	30	85	98.26388889	98.95833333	blastp
	EB656428_T1
2885	centaurea\|gb166\|	centaurea	3093	30	86	98.26388889	98.95470383	blastp
	EH767475_T1
2886	centaurea\|gb166\|	centaurea	3094	30	86	98.26388889	98.95833333	blastp
	EL935569_T1
2887	cycas\|gb166\|CB089724_T1	cycas	3095	30	84	98.26388889	98.95470383	blastp
2888	cycas\|gb166\|CB088798_T1	cycas	3096	30	83	98.26388889	98.26989619	blastp
2889	eucalyptus\|gb166\|	eucalyptus	3097	30	84	98.26388889	99.30555556	blastp
	CD668044_T1
2890	eucalyptus\|gb166\|	eucalyptus	3098	30	87	98.26388889	98.95470383	blastp
	AW191311_T1
2891	kiwi\|gb166\|FG403284_T1	kiwi	3099	30	85	98.26388889	99.3006993	blastp
2892	kiwi\|gb166\|FG404130_T1	kiwi	3100	30	86	98.26388889	99.3006993	blastp
2893	kiwi\|gb166\|FG396354_T1	kiwi	3101	30	90	98.26388889	98.95104895	blastp
2894	kiwi\|gb166\|FG404890_T1	kiwi	3102	30	85	72.22222222	99.5215311	blastp
2895	kiwi\|gb166\|FG417962_T1	kiwi	3103	30	87	62.15277778	99.44444444	blastp
2896	kiwi\|gb166\|FG403188_T1	kiwi	3104	30	89	98.26388889	96.91780822	blastp
2897	kiwi\|gb166\|FG403647_T1	kiwi	3105	30	85	68.05555556	99.49238579	blastp
2898	kiwi\|gb166\|FG397405_T1	kiwi	3106	30	85	98.26388889	99.3006993	blastp
2899	leymus\|gb166\|EG384635_T1	leymus	3107	30	85	99.30555556	99.65517241	blastp
2900	leymus\|gb166\|CN466016_T1	leymus	3108	30	83	98.26388889	98.97260274	blastp
2901	leymus\|gb166\|CN466006_T1	leymus	3109	30	83	98.26388889	98.97260274	blastp
2902	leymus\|gb166\|EG376500_T1	leymus	3110	30	83	98.26388889	98.97260274	blastp
2903	liriodendron\|gb166\|	liriodendron	3111	30	89	98.26388889	98.95470383	blastp
	CK749885_T1
2904	liriodendron\|gb166\|	liriodendron	3112	30	88	98.26388889	98.95470383	blastp
	CV002697_T1
2905	lovegrass\|gb167\|	lovegrass	3113	30	85	98.26388889	98.96193772	blastp
	DN480914_T1
2906	nuphar\|gb166\|CD472824_T1	nuphar	3114	30	86	98.26388889	98.94736842	blastp
2907	nuphar\|gb166\|CD472574_T1	nuphar	3115	30	86	98.26388889	98.94736842	blastp
2908	nuphar\|gb166\|CD473614_T1	nuphar	3116	30	84	51.73611111	98.02631579	blastp
2909	peanut\|gb167\|ES759056_T1	peanut	3117	30	83	64.23611111	100	blastp
2910	poppy\|gb166\|FE966578_T1	poppy	3118	30	83	98.61111111	98.62068966	blastp
2911	pseudoroegneria\|	pseudoroegneria	3119	30	82	98.26388889	98.97260274	blastp
	gb167\|FF344096_T1
2912	pseudoroegneria\|	pseudoroegneria	3120	30	85	98.61111111	98.62542955	blastp
	gb167\|FF346975_T1
2913	pseudoroegneria\|	pseudoroegneria	3121	30	83	98.26388889	98.97260274	blastp
	gb167\|FF342094_T1
2914	soybean\|gb167\|CA898412_T1	soybean	3122	30	84	94.44444444	95.0877193	blastp
2915	switchgrass\|gb167\|	switchgrass	3123	30	87	52.43055556	98.69281046	blastp
	FE621985_T1
2916	switchgrass\|gb167\|	switchgrass	3124	30	86	98.26388889	98.95833333	blastp
	DN141371_T1
2917	tamarix\|gb166\|CF199714_T1	tamarix	3125	30	88	78.47222222	99.12280702	blastp
2918	tamarix\|gb166\|CF226851_T1	tamarix	3126	30	85	98.26388889	99.30313589	blastp
2919	walnuts\|gb166\|CB303847_T1	walnuts	3127	30	84	98.26388889	99.31034483	blastp
2920	walnuts\|gb166\|CV194951_T1	walnuts	3128	30	86	98.26388889	99.30313589	blastp
2921	walnuts\|gb166\|CV196162_T1	walnuts	3129	30	85	98.26388889	99.31506849	blastp
2922	zamia\|gb166\|FD765004_T1	zamia	3130	30	84	98.26388889	98.95470383	blastp
2923	antirrhinum\|gb166\|	antirrhinum	3131	31	80	98.7854251	100	blastp
	AJ799752_T1
2924	kiwi\|gb166\|FG400670_T1	kiwi	3132	31	82	74.08906883	100	blastp
2925	kiwi\|gb166\|FG418275_T1	kiwi	3133	31	83	98.7854251	98.38709677	blastp
2926	centaurea\|gb166\|	centaurea	3134	34	83	88.57142857	32.74021352	blastp
	EL931588_T1
2927	soybean\|gb167\|AW119586_T1	soybean	3135	34	83	94.28571429	36.26373626	blastp
2928	soybean\|gb167\|AW573764_T1	soybean	3136	34	83	94.28571429	36.66666667	blastp
2929	amborella\|gb166\|	amborella	3137	35	81	65.76271186	100	blastp
	FD440187_T1
2930	centaurea\|gb166\|	centaurea	3138	35	82	97.62711864	96.61016949	blastp
	EL931433_T1
2931	eucalyptus\|gb166\|	eucalyptus	3139	35	85	73.89830508	100	blastp
	CD668486_T1
2932	petunia\|gb166\|DC243166_T1	petunia	3140	36	86	100	100	blastp
2933	amborella\|gb166\|	amborella	3141	39	82	99.64157706	98.93992933	blastp
	CD482946_T1
2934	antirrhinum\|gb166\|	antirrhinum	3142	39	88	99.28315412	98.57651246	blastp
	AJ559435_T1
2935	antirrhinum\|gb166\|	antirrhinum	3143	39	89	71.32616487	100	blastp
	AJ793990_T1
2936	antirrhinum\|gb166\|	antirrhinum	3144	39	83	61.29032258	87.24489796	blastp
	AJ559760_T1
2937	antirrhinum\|gb166\|	antirrhinum	3145	39	88	88.53046595	100	blastp
	AJ558545_T1
2938	bean\|gb167\|FE682762_T1	bean	3146	39	86	100	100	blastp
2939	bean\|gb167\|CV531088_T1	bean	3147	39	86	99.64157706	98.94736842	blastp
2940	bean\|gb167\|CA907460_T1	bean	3148	39	86	99.64157706	98.94736842	blastp
2941	cenchrus\|gb166\|	cenchrus	3149	39	83	97.49103943	95.86206897	blastp
	EB655519_T1
2942	centaurea\|gb166\|	centaurea	3150	39	88	53.76344086	96.15384615	blastp
	EL934360_T1
2943	centaurea\|gb166\|	centaurea	3151	39	80	88.53046595	100	blastp
	EH751120_T1
2944	centaurea\|gb166\|	centaurea	3152	39	87	83.5125448	100	blastp
	EH752971_T1
2945	centaurea\|gb166\|	centaurea	3153	39	80	91.7562724	99.61685824	blastp
	EH768434_T1
2946	centaurea\|gb166\|	centaurea	3154	39	83	83.15412186	100	blastp
	EH767287_T1
2947	cichorium\|gb166\|	cichorium	3155	39	82	89.96415771	100	blastp
	DT212008_T1
2948	cichorium\|gb166\|	cichorium	3156	39	84	86.73835125	97.61904762	blastp
	EL354583_T1
2949	eucalyptus\|gb166\|	eucalyptus	3157	39	87	83.87096774	100	blastp
	CD668553_T1
2950	eucalyptus\|gb166\|	eucalyptus	3158	39	80	69.89247312	100	blastp
	CD668534_T1
2951	eucalyptus\|gb166\|	eucalyptus	3159	39	88	100	99.3006993	blastp
	CD668523_T1
2952	eucalyptus\|gb166\|	eucalyptus	3160	39	83	100	100	blastp
	CD669942_T1
2953	kiwi\|gb166\|FG405216_T1	kiwi	3161	39	84	100	99.29328622	blastp
2954	kiwi\|gb166\|FG495821_T1	kiwi	3162	39	86	50.17921147	100	blastp
2955	kiwi\|gb166\|FG417997_T1	kiwi	3163	39	86	64.51612903	100	blastp
2956	kiwi\|gb166\|FG403725_T1	kiwi	3164	39	83	73.11827957	100	blastp
2957	kiwi\|gb166\|FG397310_T1	kiwi	3165	39	88	100	100	blastp
2958	kiwi\|gb166\|FG408531_T1	kiwi	3166	39	83	100	99.30313589	blastp
2959	leymus\|gb166\|EG381236_T1	leymus	3167	39	81	55.55555556	97.46835443	blastp
2960	leymus\|gb166\|EG376087_T1	leymus	3168	39	80	96.77419355	96.55172414	blastp
2961	leymus\|gb166\|CD808804_T1	leymus	3169	39	81	100	100	blastp
2962	leymus\|gb166\|EG378918_T1	leymus	3170	39	86	51.25448029	100	blastp
2963	liriodendron\|gb166\|	liriodendron	3171	39	81	99.64157706	98.96193772	blastp
	CK761396_T1
2964	nuphar\|gb166\|ES730700_T1	nuphar	3172	39	81	61.29032258	98.27586207	blastp
2965	poppy\|gb166\|FE965621_T1	poppy	3173	39	84	88.53046595	100	blastp
2966	pseudoroegneria\|	pseudoroegneria	3174	39	81	100	100	blastp
	gb167\|FF340233_T1
2967	switchgrass\|gb167\|	switchgrass	3175	39	80	100	100	blastp
	FE638368_T1
2968	switchgrass\|gb167\|	switchgrass	3176	39	80	99.28315412	98.95833333	blastp
	FE657460_T1
2969	switchgrass\|gb167\|	switchgrass	3177	39	82	78.85304659	86.59003831	blastp
	FE641178_T1
2970	switchgrass\|gb167\|	switchgrass	3178	39	82	94.26523297	95.72953737	blastp
	FE619224_T1
2971	switchgrass\|gb167\|	switchgrass	3179	39	81	100	100	blastp
	FE657460_T2
2972	tamarix\|gb166\|EH051524_T1	tamarix	3180	39	82	56.27240143	98.74213836	blastp
2973	walnuts\|gb166\|CB304207_T1	walnuts	3181	39	84	100	99.30313589	blastp
2974	walnuts\|gb166\|CB303561_T1	walnuts	3182	39	84	100	100	blastp
2975	walnuts\|gb166\|EL892579_T1	walnuts	3183	39	84	89.60573477	98.82352941	blastp
2976	zamia\|gb166\|DY032141_T1	zamia	3184	39	80	96.05734767	94.32624113	blastp
2977	zamia\|gb166\|FD764669_T1	zamia	3185	39	80	99.64157706	98.92473118	blastp
2978	zamia\|gb166\|DY034152_T1	zamia	3186	39	80	94.62365591	92.25352113	blastp
2979	antirrhinum\|gb166\|	antirrhinum	3187	40	87	100	100	blastp
	AJ568195_T1
2980	antirrhinum\|gb166\|	antirrhinum	3188	40	87	100	100	blastp
	AJ568110_T1
2981	banana\|gb167\|FL657842_T1	banana	3189	40	81	83.03886926	92.49011858	blastp
2982	bruguiera\|gb166\|	bruguiera	3190	40	84	100	100	blastp
	EF126757_T1
2983	centaurea\|gb166\|	centaurea	3191	40	84	92.5795053	100	blastp
	EH765776_T1
2984	eucalyptus\|gb166\|	eucalyptus	3192	40	85	100	100	blastp
	AJ627837_T1
2985	kiwi\|gb166\|FG411924_T1	kiwi	3193	40	86	100	100	blastp
2986	kiwi\|gb166\|FG400706_T1	kiwi	3194	40	85	100	100	blastp
2987	kiwi\|gb166\|FG418898_T1	kiwi	3195	40	84	71.02473498	98.5	blastp
2988	kiwi\|gb166\|FG420187_T1	kiwi	3196	40	85	72.79151943	100	blastp
2989	kiwi\|gb166\|FG398010_T1	kiwi	3197	40	86	100	100	blastp
2990	petunia\|gb166\|AF452012_T1	petunia	3198	40	88	100	100	blastp
2991	tamarix\|gb166\|CV121772_T1	tamarix	3199	40	83	100	100	blastp
2992	walnuts\|gb166\|EL893208_T1	walnuts	3200	40	84	100	100	blastp
2993	bean\|gb167\|FD786218_T1	bean	3201	41	83	58.60927152	100	blastp
2994	citrus\|gb166\|CX074333_T2	citrus	3202	41	81	72.51655629	99.5412844	blastp
2995	citrus\|gb166\|CX074333_T1	citrus	3203	41	83	96.02649007	91.42857143	blastp
2996	kiwi\|gb166\|FG420468_T2	kiwi	3204	41	84	87.08609272	97.04797048	blastp
2997	kiwi\|gb166\|FG420468_T1	kiwi	3205	41	83	58.94039735	95.69892473	blastp
2998	tamarix\|gb166\|EG972096_T1	tamarix	3206	42	82	60.97560976	95.23809524	blastp
2999	pseudoroegneria\|	pseudoroegneria	3207	43	86	98.3935743	98.79518072	blastp
	gb167\|FF353501_T1
3000	switchgrass\|gb167\|	switchgrass	3208	43	86	98.79518072	99.19678715	blastp
	FE646459_T1
3001	switchgrass\|gb167\|	switchgrass	3209	43	84	71.48594378	93.22916667	blastp
	FL887003_T1
3002	wheat\|gb164\|BE403397_T1	wheat	3210	43	86	98.3935743	98.79518072	blastp
3003	leymus\|gb166\|EG381168_T1	leymus	3211	44	96	52.01612903	100	blastp
3004	pseudoroegneria\|	pseudoroegneria	3212	44	97	100	100	blastp
	gb167\|FF341567_T1
3005	switchgrass\|gb167\|	switchgrass	3213	44	91	100	100	blastp
	FE618822_T1
3006	switchgrass\|gb167\|	switchgrass	3214	44	92	78.22580645	100	blastp
	FE633786_T1
3007	eucalyptus\|gb166\|	eucalyptus	3215	46	84	97.51552795	60.78431373	blastp
	CB967586_T1
3008	liriodendron\|gb166\|	liriodendron	3216	46	80	74.53416149	100	blastp
	CK762443_T1
3009	switchgrass\|gb167\|	switchgrass	3217	46	90	100	61.38996139	blastp
	FE615387_T1
3010	walnuts\|gb166\|EL893973_T1	walnuts	3218	46	82	97.51552795	55.47445255	blastp
3011	bean\|gb167\|FD782805_T1	bean	3219	47	91	88.17204301	79.61165049	blastp
3012	bean\|gb167\|EY457935_T1	bean	3220	47	82	100	38.58921162	blastp
3013	cichorium\|gb166\|	cichorium	3221	47	80	100	67.39130435	blastp
	EH685648_T2
3014	cichorium\|gb166\|	cichorium	3222	47	80	100	32.1799308	blastp
	EH685648_T1
3015	citrus\|gb166\|CK701147_T1	citrus	3223	47	90	100	76.85950413	blastp
3016	citrus\|gb166\|CX544905_T1	citrus	3224	47	81	79.56989247	88.0952381	blastp
3017	cycas\|gb166\|CB088978_T1	cycas	3225	47	82	87.09677419	30.68181818	blastp
3018	cycas\|gb166\|EX920749_T1	cycas	3226	47	88	100	76.2295082	blastp
3019	kiwi\|gb166\|FG429765_T1	kiwi	3227	47	84	98.92473118	77.96610169	blastp
3020	leymus\|gb166\|CN466394_T1	leymus	3228	47	87	100	79.48717949	blastp
3021	leymus\|gb166\|EG376019_T1	leymus	3229	47	88	100	32.74647887	blastp
3022	leymus\|gb166\|EG390723_T1	leymus	3230	47	81	100	31.95876289	blastp
3023	leymus\|gb166\|EG387193_T1	leymus	3231	47	81	100	32.1799308	blastp
3024	nuphar\|gb166\|CD473277_T1	nuphar	3232	47	91	100	75.6097561	blastp
3025	nuphar\|gb166\|FD384794_T1	nuphar	3233	47	86	96.77419355	81.08108108	blastp
3026	nuphar\|gb166\|CD475538_T1	nuphar	3234	47	89	100	69.92481203	blastp
3027	nuphar\|gb166\|CD472711_T1	nuphar	3235	47	91	100	48.94736842	blastp
3028	petunia\|gb166\|DC240378_T1	petunia	3236	47	92	58.06451613	98.18181818	blastp
3029	pseudoroegneria\|	pseudoroegneria	3237	47	87	100	32.74647887	blastp
	gb167\|FF344283_T1
3030	sorghum\|gb161.crp\|	sorghum	3238	47	92	100	32.51748252	blastp
	AI724931_T1
3031	switchgrass\|gb167\|	switchgrass	3239	47	82	100	33.69565217	blastp
	FL923354_T1
3032	switchgrass\|gb167\|	switchgrass	3240	47	92	98.92473118	74.79674797	blastp
	FE657461_T1
3033	switchgrass\|gb167\|	switchgrass	3241	47	82	100	72.22222222	blastp
	FL765830_T1
3034	switchgrass\|gb167\|	switchgrass	3242	47	83	97.84946237	87.5	blastp
	FL915169_T1
3035	tamarix\|gb166\|EH054604_T1	tamarix	3243	47	83	100	65.64705882	tblastn
3036	walnuts\|gb166\|CB303798_T1	walnuts	3244	47	91	100	80.17241379	blastp
3037	bruguiera\|gb166\|	bruguiera	3245	48	82	52.05479452	100	blastp
	BP942548_T1
3038	bruguiera\|gb166\|	bruguiera	3246	48	89	56.62100457	91.17647059	blastp
	BP938825_T1
3039	centaurea\|gb166\|	centaurea	3247	48	83	87.21461187	50	tblastn
	EH739326_T1
3040	eucalyptus\|gb166\|	eucalyptus	3248	48	87	56.16438356	91.79104478	blastp
	CD669176_T1
3041	leymus\|gb166\|EG382428_T1	leymus	3249	48	80	52.05479452	90.47619048	blastp
3042	liriodendron\|gb166\|	liriodendron	3250	48	81	94.06392694	54.54545455	tblastn
	CK743477_T1
3043	marchantia\|gb166\|	marchantia	3251	48	83	94.06392694	72.53521127	blastp
	BJ840587_T1
3044	marchantia\|gb166\|	marchantia	3252	48	83	93.15068493	71.57894737	blastp
	C96070_T1
3045	nuphar\|gb166\|CK748374_T1	nuphar	3253	48	82	65.75342466	99.31034483	blastp
3046	pseudoroegneria\|	pseudoroegneria	3254	48	81	96.80365297	73.44827586	blastp
	gb167\|FF340047_T1
3047	pseudoroegneria\|	pseudoroegneria	3255	48	81	96.80365297	73.44827586	blastp
	gb167\|FF340899_T1
3048	pseudoroegneria\|	pseudoroegneria	3256	48	82	96.80365297	74.12587413	blastp
	gb167\|FF352644_T1
3049	sorghum\|gb161.crp\|	sorghum	3257	48	80	96.80365297	74.12587413	blastp
	SBGWP030188_T1
3050	switchgrass\|gb167\|	switchgrass	3258	48	81	96.80365297	74.12587413	blastp
	FL718379_T1
3051	switchgrass\|gb167\|	switchgrass	3259	48	82	96.80365297	73.10344828	blastp
	FE639195_T1

Table 3: Homologues and orthologues of the AQP proteins are provided.
Homology was calculated as % of identity over the aligned sequences.
Polynuc. = Polynucleotide;
Polypep. = Polypeptide;
Hom. = Homologues/Orthologues;
% Ident. = percent identity;
Cover. = coverage.

Example 2

mRNA Expression of In-Silico Expressed Polynucleotides

Messenger RNA levels were determined using reverse transcription assay followed by quantitative Real-Time PCR (qRT-PCR) analysis. RNA levels were compared between leaves of 20 days old seedlings of tomato plants grown under salinity water. A correlation analysis between mRNA levels in different experimental conditions/genetic backgrounds was performed in order to determine the role of the gene in the plant.
Materials and Experimental Methods
Quantitative Real Time RT-PCR (qRT-PCR)—To verify the level of expression, specificity and trait-association, Reverse Transcription followed by quantitative Real-Time PCR (qRTPCR) was performed on total RNA extracted from leaves of 2 tomato varieties namely YO361 (salt tolerant variety) and FA191 (salt sensitive variety). Messenger RNA (mRNA) levels were determined for AQP genes, expressed under normal and stressed conditions.
Twenty days-old tomato seedlings were grown in soil and soaked with 300 mM NaCl for 0, 1, 6, 24, 118 hours. Leaves were harvested and frozen in liquid nitrogen and then kept at −80° C. until RNA extraction. Total RNA was extracted from leaves using RNeasy plant mini kit (Qiagen, Hilden, Germany) and by using the protocol provided by the manufacturer. Reverse transcription was performed using 1 μg total RNA, using 200 U Super Script II Reverse Transcriptase enzyme (Invitrogen), 150 ng random deoxynucleotide hexamers (Invitrogen), 500 μM deoxynucleotide tri-phosphates (dNTPs) mix (Takara, Japan), 0.2 volume of ×5 reverse transcriptase (RT) buffer (Invitrogen), 0.01 M dithiothreitol (DTT), 40 U RNAsin (Promega), diethylpyrocarbonate (DEPC) treated double distilled water (DDW) was added up to 24 μl.
Mix of RNA, random deoxynucleotide hexamers, dNTPs mix and DEPC treated DDW was incubated at 65° C. for 5 minutes, followed by 4° C. for 5 minutes. Mix of reverse transcriptase (RT) buffer, dithiothreitol (DTT) and RNAsin was added to the RT reactions followed by incubation at 25° C. for 10 minutes and at 42° C. for 2 minutes afterwards. Finally, Super Script II Reverse Transcriptase enzyme was added to the RT reactions that were further incubated for 50 minutes at 42° C., followed by 70° C. for 15 minutes.
cDNA was diluted 1:20 in Tris EDTA, pH=7.5 for MAB69 and housekeeping genes. For MAB58 and MAB59 cDNA was diluted 1:2 due to very weak expression and consequently for housekeeping genes cDNA was diluted 1:8 in order to insert the Ct values in calibration curve range. 5 μL of the diluted cDNA was used for qPCR.
For qPCR amplification, primers of the AQP genes were designed, as summarized in Table 4 below. The expression level of the housekeeping genes: Actin (SEQ ID NO: 2841), GAPDH (SEQ ID NO: 2842) and RPL19 (SEQ ID NO: 2747) was determined in order to normalize the expression level between the different tissues.

TABLE 4

Table 4
Primers for qPCR amplification

	Forward	Forward	Reverse	Reverse
	primer	primer	primer	primer
	SEQ ID	sequence	SEQ ID	sequence
Gene	NO:	(5′→3′)	NO:	(5′→3′)

MAB58	2829	CTTTTGGTAGG	2830	CGAAGATGAAGG
		GCCGATGAAG		TGGATAAGAGCT

MAB59	2831	CAGTATGAACG	2832	CAACAGCACCT
		TCTCCGGTGG		AGCAACTGACC

MAB69	2833	TGTCTTGGATTC	2834	GTTTGAGCTG
		CATTGAGCACT		CTGTCCCCA

Actin	2835	CCACATGCCA	2836	GCTTTTCTTTC
(SEQ		TTCTCCGTCT		ACGTCCCTGA
ID NO:
2841)

GAPD	2837	TTGTTGTGGGT	2838	ATGGCGTGGA
H (SEQ		GTCAACGAGA		CAGTGGTCA
ID NO:
2842)

RPL19	2839	CACTCTGGATATG	2840	TTCTTGGACTCC
(SEQ		GTAAGCGTAAGG		CTGTACTTACGA
ID NO:
2747)

Experimental Results
Changes in mRNA levels of AQP genes in leaves of plants under salt tolerance—Steady state levels of tomato AQP genes in leaves of tolerant versus sensitive lines, under salinity conditions are summarized in Table 5 below. In all 3 cases, aquaporin gene expression was increased after plant was exposed to salt stress. Gene peak expression was higher in the salt tolerance tomato line (YO361) versus the sensitive line (FA191). The elevated gene expression demonstrates the involvement of the tested AQP genes in tomato plants tolerating high salinity.

TABLE 5

Expression levels of tomato AQP genes

	Well name (cDNA)	MAB58	MAB59	MAB69*

leaf FA191 0 h	2.86	5.97	875
leaf FA191 1 h	4.56	5.73	597
leaf FA191 6 h	5.34	8.2	945
leaf FA191 24 h	42.7	62.4	613
leaf FA191 118 h	55.4	22.2	1800
leaf Y0361 0 h	1.96	2.81	638
leaf Y0361 1 h	2.33	0.517	583
leaf Y0361 6 h	2.88	5.66	464
leaf Y0361 24 h	75.4	139	513
leaf Y0361 118 h	26.3	Not determined	2300

Table 5: Provided are the steady state levels of tomato AQP genes under salinity conditions [the incubation periods in the salt solution are provided in hours (h)]. Different dilutions of cDNA were used (1:20 for MAB69 and 1:2 for MAB58 and MAB59). Numbers are given after normalization for each sample.

Example 3

Gene Cloning and Generation of Binary Vectors for Plant Expression

To validate their role in improving ABST and yield, the AQP genes were over-expressed in plants, as follows.
Cloning Strategy
Selected genes from those presented in Example 1 were cloned into binary vectors for the generation of transgenic plants. For cloning, the full-length open reading frames (ORFs) were identified. EST clusters and in some cases mRNA sequences were analyzed to identify the entire open reading frame by comparing the results of several translation algorithms to known proteins from other plant species. In case where the entire coding sequence is not found, RACE kits from Ambion or Clontech (RACE=Rapid Access to cDNA Ends) were used to prepare RNA from the plant samples to thereby access the full cDNA transcript of the gene.
In order to clone the full-length cDNAs, Reverse Transcription followed by PCR (RT-PCR) was performed on total RNA extracted from leaves, roots or other plant tissues, growing under either normal or nutrient deficient conditions. Total RNA extraction, production of cDNA and PCR amplification was performed using standard protocols described elsewhere (Sambrook J., E. F. Fritsch, and T. Maniatis. 1989. Molecular Cloning. A Laboratory Manual., 2nd Ed. Cold Spring Harbor Laboratory Press, New York.), and are basic for those skilled in the art. PCR products were purified using PCR purification kit (Qiagen) and sequencing of the amplified PCR products was performed, using ABI 377 sequencer (Applied Biosystems).
Usually, 2 sets of primers were ordered for the amplification of each gene, via nested PCR (meaning first amplifying the gene using external primers and then using the produced PCR product as a template for a second PCR reaction, where the internal set of primers are used). Alternatively, one or two of the internal primers were used for gene amplification, both in the first and the second PCR reactions (meaning only 2-3 primers were designed for a gene). To facilitate further cloning of the cDNAs, a 8-12 bp extension is added to the 5′ primer end of each internal primer. The primer extension includes an endonuclease restriction site. The restriction sites are selected using two parameters: (a) The restriction site does not exist in the cDNA sequence; and (b) The restriction sites in the forward and reverse primers are designed so the digested cDNA is inserted in the sense formation into the binary vector utilized for transformation. In Table 6 below, primers used for cloning tomato and barley AQPs are provided.

TABLE 6

Table 6
Primers used for cloning tomato and barley AQP genes

	Forward external	Forward internal	Reverse external	Reverse internal
MAB	primer sequence	primer sequence	primer sequence	primer sequence
gene	from 5′→3′	from 5′→3′	from 5′→3′	from 5′→3′
No.	(SEQ ID NO:)	(SEQ ID NO:)	(SEQ ID NO:)	(SEQ ID NO:)

55	GGAGTCGACGACCAT	GGAGTCGACTT	TGAGCTCACTTC	TGAGCTCCCATCC
	CAAGTTTTAAGTGAC	AAGTACATTCTT	AAAACCATCCG	GTTGTCAAAATGA
	TTC (2770)	TAGTGAGAGCC	TTGTC (2772)	AC (2773)
		(2771)

56		GGAGTCGACGT	CGAGCTCGTAA	CGAGCTCAAGAC
		AAGAAACAATA	AGCCAAGTTTTG	AAACAAAGAGAA
		ATGCCAATTTC	AAAGAC (2775)	GAGGG (2776)
		(2774)

57		GTTAAAAATGC	GCGATATCTAA	GCGATATCAGCCG
		CGATCAACC	ATAACAAAAGC	TCCGAATAAACAA
		(2777)	CGTCCG (2778)	AG (2779)

58	AATGTCGACCGAATT	TATGTCGACTTC	TTTCTAGAGGTC	TTTCTAGAGATGT
	GATCTCCTTCTTGATC	ATTTCTTGGGTC	TGGGATTATCGT	GCAGGCAGCTAC
	(2780)	ACTCG (2781)	CTTG (2782)	ATAC (2783)

59	AATGTCGACTTTAAG	AATGTCGACTC		AATCTAGATTAGA
	CGGTGTGTTTTGTG	ACAATTATGCA		CCCAAACATACAA
	(2784)	GCCACG (2785)		ACTTCAC (2786))

69	TAAGTCGACACAAAC	AATGTCGACCTT		TGAGCTCTGGAGA
	CTTATCCTGGTCTCAT	GGATTCCATTGA		AAGAAAACTTTAG
	C (2787)	GCACTC (2788)		ATACA (2789)

70	AACTGCAGAGCTGTA	AACTGCAGTGT	TCCCGGGCCAG	TCCCGGGCTTCAA
	CATGGTCCTCCTCC	ACATGGTCCTCC	ACAAAACTTCA	TTTCATCTTCTGA
	(2790)	TCCG (2791)	ATTTCATC (2792)	TTTC (2793)

71	AAAGTCGACGGAAAA	TTTGTCGACCTT	AATCTAGACAA	AATCTAGAGTACT
	TGCATTAAAACCTTA	AGTTTTCTCCCA	GTAGAGGTACT	AGGTAGGGACAA
	AG (2794)	CATATGG (2795)	AGGTAGGGAC	TATGATATG
			(2796)	(2797)

72	AATGTCGACGTGGAG	AATGTCGACCTC		TATCTAGAGGATG
	GAGGAGTCTTTGATA	CAACACTCTTAT		CAACTACAAAGA
	C (2798)	CAATTACCA		AATTG (2800)
		(2799)

74		TTCTGCAGGTTT		TCCCGGGGCATAG
		GGGAGTTATTG		TTCACACAGAGCA
		ATCTAAGATG		AATC (2802)
		(2801)

76	AATGTCGACCTGTAT	AATGTCGACGT	TTTCTAGACTAG	AATCTAGATTAGA
	CCTCTTAAGTATGAA	CGTCTTGTATGT	TGGTATAGATC	GCTGGAGAATGA
	TCG (2803)	ATTTGTACTACT	ATTTTATGGTGA	ACTGAAGC (2806)
		G (2804)	C (2805)

77	AACTGCAGCTTCTTTC	AACTGCAGCTTT	ACCCGGGAATT	TCCCGGGTCCAAC
	ACCGAGTGGGAG	CACCGAGTGGG	CCAACTAGCTGT	TAGCTGTTATGAT
	(2807)	AGAG (2808)	TATGATTC	TCTG (2810)
			(2809)

79	AAGATATCAAAAAAA	AAGATATCAAC		AAGATATCGACCA
	TGTCGAAGGACGTG	AATGTCGAAGG		CCAACTCTAGTCT
	(2811)	ACGTGATTGA		CATACC (2813)
		(2812)

115	AGATTCGAATCTTTA	AATCTAGAGAA		AGAGCTCTTAAGG
	GCCTG	GTCACAGAGAA		GAAATTCATCACA
	(2814)	AACAGTCGAG		CAAGG (2816)
		(2815)

116	AAAGTCGACCTCATC	TTTGTCGACCAT	TATCTAGAATTG	TTTCTAGAGACCG
	AGTGTTAAAGCCATA	AAGCCCTCTTTG	AATCGAAAGGG	TGACACACCATTT
	AG (2817)	AGTGTG (2818)	AAACAC (2819)	GTAC (2820)

117		TTTCTAGACTCA		TGAGCTCCAGATA
		GCGACAACATT		GAGAAGCATGCA
		TCATCTC (2821)		TCATC (2822)

PCR products were purified (PCR Purification Kit, Qiagen, Germany) and digested with the restriction endonucleases (Roche, Switzerland) according to the sites design in the primers (Tables 8 and 9 below). Each of the digested PCR products was cloned first into high copy plasmid pBlue-script KS [Hypertext Transfer Protocol://World Wide Web (dot) stratagene (dot) com/manuals/212205 (dot) pdf] which was digested with the same restriction enzymes. In some cases (Table 8, below) the Nopaline Synthase (NOS) terminator originated from the binary vector pBI101.3 [nucleotide coordinates 4417-4693 in GenBank Accession No. U12640 (SEQ ID NO:2824)] was already cloned into the pBlue-script KS, between the restriction endonuclease sites SacI and EcoRI, so the gene is introduced upstream of the terminator. In other cases (Table 9, below) the At6669 promoter (SEQ ID NO: 2823) is already cloned into the pBlue-script KS, so the gene is introduced downstream of the promoter. The digested PCR products and the linearized plasmid vector were ligated using T4 DNA ligase enzyme (Roche, Switzerland). Sequencing of the inserted genes was conducted, using ABI 377 sequencer (Applied Biosystems). Sequences of few of the cloned AQP genes, as well as their encoded proteins are listed in Table 7, below.

TABLE 7

Cloned sequences

Serial		Polynucleotide	Polypeptide
No	Gene Name	SEQ ID NO:	SEQ ID NO:

1	MAB115	2748	2765
2	MAB116	2749	47
3	MAB117	2750	48
4	MAB54	2751	27
5	MAB55	2752	28
6	MAB56	2753	29
7	MAB57	2754	30
8	MAB58	2755	2766
9	MAB59	2756	2767
10	MAB69	2757	33
11	MAB70	2758	34
12	MAB71	2759	2768
13	MAB72	2760	2769
14	MAB72	2843	2769
	(Optimized for
	expression in Arabidopsis,
	Tomato and Maize)
15	MAB74	2761	38
16	MAB76	2762	40
17	MAB77	2763	41
18	MAB79	2764	43

Table 7.

The genes were digested again and ligated into pPI or pGI binary plasmids, harboring the At6669 promoter (between the HindIII and SalI restriction endonucleases site) (Table 8). In other cases the At6669 promoter together with the gene are digested out of the pBlue-script KS plasmid and ligated into pPI or pGI binary plasmids, using restriction endonucleases as given in Table 8.

TABLE 8

Restriction enzyme sites used to clone the MAB AQP genes into pKS + NOS terminator
high copy plasmid, followed by cloning into the binary vector pGI + At6669 promoter

	Restriction enzymes	Restriction enzymes	Restriction enzymes	Restriction enzymes
	used for cloning	used for cloning	used for cloning	used for cloning	Restriction enzymes
MAB gene No.	into high copy	into high copy	into binary vector-	into binary vector-	used for digesting
(SEQ ID NO:)	plasmid-FORWARD	plasmid-REVERSE	FORWARD	REVERSE	the binary vector

54 (SEQ	XbaI	SacI	SalI	EcoRI	SalI/EcoRI
ID NO: 2751)
55 (SEQ	SalI	SacI	SalI	EcoRI	SalI/EcoRI
ID NO: 2752)
56 (SEQ	SalI	SacI	SalI	EcoRI	SalI/EcoRI
ID NO: 2753)
58 (SEQ	SalI	XbaI	SalI	EcoRI	SalI/EcoRI
ID NO: 2755)
59 (SEQ	SalI	XbaI	SalI	EcoRI	SalI/EcoRI
ID NO: 2756)
69 (SEQ	SalI	SacI	SalI	EcoRI	SalI/EcoRI
ID NO: 2757)
71 (SEQ	SalI	XbaI	SalI	EcoRI	SalI/EcoRI
ID NO: 2759)
72 (SEQ	SalI	XbaI	SalI	EcoRI	SalI/EcoRI
ID NO: 2760)
76 (SEQ	SalI	XbaI	SalI	EcoRI	SalI/EcoRI
ID NO: 2762)
115 (SEQ	XbaI	SacI	SalI	EcoRI	SalI/EcoRI
ID NO: 2748)
116 (SEQ	SalI	XbaI	SalI	EcoRI	SalI/EcoRI
ID NO: 2749)
117 (SEQ	XbaI	SacI	SalI	EcoRI	SalI/EcoRI
ID NO: 2750)

Table 8: MAB AQP genes cloned into pKS + NOS terminator high copy plasmid, followed by cloning into the binary vector pGI + At6669 promoter.

TABLE 9

Restriction enzyme sites used to clone the MAB AQP genes into pKS + At6669
promoter high copy plasmid, followed by cloning promoter + gene into pGI binary vector

	Restriction enzymes	Restriction enzymes	Restriction enzymes	Restriction enzymes
	used for cloning	used for cloning	used for cloning	used for cloning	Restriction enzymes
MAB gene No.	into high copy	into high copy	into binary vector-	into binary vector-	used for digesting
(SEQ ID NO:)	plasmid-FORWARD	plasmid-REVERSE	FORWARD	REVERSE	the binary vector

57 (SEQ	Blunt	EcoRV	SalI	EcoRV	SalI/Ecl136 II
ID NO: 2754)
70 (SEQ	PstI	SmaI	BamHI	SmaI	BamHI/Ecl136II
ID NO: 2758)
74 (SEQ	PstI	SmaI	BamHI	SmaI	BamHI/Ecl136II
ID NO: 2761)
77 (SEQ	PstI	SmaI	BamHI	SmaI	BamHI/Ecl136II
ID NO: 2763)
79 (SEQ	EcoRI	EcoRV	SalI	SmaI	SalI/Ecl136II
ID NO: 2764)

Table 9: MAB AQP genes cloned into pKS + At6669 promoter high copy plasmid, followed by cloning promoter + gene into pGI binary vector.

The pPI plasmid vector was constructed by inserting a synthetic poly-(A) signal sequence, originating from pGL3 basic plasmid vector (Promega, Acc. No. U47295; by 4658-4811) into the HindIII restriction site of the binary vector pBI101.3 (Clontech, Acc. No. U12640). pGI (FIG. 1) is similar to pPI, but the original gene in the back bone is GUS-Intron, rather than GUS. The cloned genes were sequenced.
Synthetic sequences (such as of MAB54, nucleotide SEQ ID NO: 2751, which encodes protein SEQ ID NO: 27; or MAB72 SEQ ID NO:2843, which encodes SEQ ID NO:2769) of some of the cloned polynucleotides were ordered from a commercial supplier (GeneArt, GmbH). To optimize the coding sequence, codon-usage Tables calculated from plant transcriptomes were used [example of such Tables can be found in the Codon Usage Database available online at Hypertext Transfer Protocol://World Wide Web (dot) kazusa (dot) or (dot) jp/codon/]. The optimized coding sequences were designed in a way that no changes were introduced in the encoded amino acid sequence while using codons preferred for expression in dicotyledonous plants mainly tomato and Arabidopsis; and monocotyledonous plants such as maize. Such optimized sequences promote better translation rate and therefore higher protein expression levels. To the optimized sequences flanking additional unique restriction enzymes sites were added to facilitate cloning genes in binary vectors.
Promoters used: CaMV 35S promoter (SEQ ID NO: 2825) and Arabidopsis At6669 promoter (SEQ ID NO: 2823; which is SEQ ID NO:61 of WO04081173 to Evogene Ltd.).

Example 4

Generation of Transgenic Plants Expressing the AQP Genes

Experimental Results
Arabidopsis transformation—Arabidopsis transformation of the following MAB genes and orthologues: MAB115, MAB54, MAB55, MAB56, MAB57, MAB58, MAB59 (ortholog of MAB58), MAB69, MAB70, MAB71, MAB72, MAB74, MAB76, MAB77, MAB79, MAB116 (ortholog of MAB115 and MAB55), and MAB117 (the sequence identifiers of the cloned polynucleotides and their expressed polypeptides are provided in Table 7 above) was performed according to Clough S J, Bent A F. (1998) “Floral dip: a simplified method for Agrobacterium-mediated transformation of Arabidopsis thaliana.” Plant J. 16(6): 735-43; and Desfeux C, Clough S J, Bent A F. (2000) “Female reproductive tissues are the primary targets of Agrobacterium-mediated transformation by the Arabidopsis floral-dip method.” Plant Physiol. 123(3): 895-904; with minor modifications. Briefly, Arabidopsis thaliana Columbia (Co10) T₀Plants were sown in 250 ml pots filled with wet peat-based growth mix. The pots were covered with aluminum foil and a plastic dome, kept at 4° C. for 3-4 days, then uncovered and incubated in a growth chamber at 18-24° C. under 16/8 hours light/dark cycles. The T₀plants were ready for transformation six days prior to anthesis. Single colonies of Agrobacterium carrying the binary vectors harboring the AQP genes are cultured in LB medium supplemented with kanamycin (50 mg/L) and gentamycin (50 mg/L). The cultures were incubated at 28° C. for 48 hours under vigorous shaking and centrifuged at 4000 rpm for 5 minutes. The pellets comprising Agrobacterium cells were resuspended in a transformation medium which contained half-strength (2.15 g/L) Murashige-Skoog (Duchefa); 0.044 μM benzylamino purine (Sigma); 112 μg/L B5 Gambourg vitamins (Sigma); 5% sucrose; and 0.2 ml/L Silwet L-77 (OSI Specialists, CT) in double-distilled water, at pH of 5.7.
Transformation of T₀plants was performed by inverting each plant into an Agrobacterium suspension such that the flowering stem was submerged for 3-5 seconds. Each inoculated T₀plant was immediately placed in a plastic tray, then covered with clear plastic dome to maintain humidity and kept in the dark at room temperature for 18 hours to facilitate infection and transformation. Transformed (transgenic) plants were then uncovered and transferred to a greenhouse for recovery and maturation. The transgenic T₀plants were grown in the greenhouse for 3-5 weeks until siliques maturation, and then seeds were harvested and kept at room temperature until sowing.
For generating T1 and T₂transgenic plants harboring the genes, seeds collected from transgenic T₀plants are surface-sterilized by soaking in 70% ethanol for 1 minute, followed by soaking in 5% sodium hypochlorite and 0.05% Triton X-100 for 5 minutes. The surface-sterilized seeds are thoroughly washed in sterile distilled water then placed on culture plates containing half-strength Murashige-Skoog (Duchefa); 2% sucrose; 0.8% plant agar; 50 mM kanamycin; and 200 mM carbenicylin (Duchefa). The culture plates are incubated at 4° C. for 48 hours then transferred to a growth room at 25° C. for an additional week of incubation. Vital T₁ Arabidopsis plants are transferred to fresh culture plates for another week of incubation. Following incubation the T₁plants are removed from culture plates and planted in growth mix contained in 250 ml pots. The transgenic plants were allowed to grow in a greenhouse to maturity. Seeds harvested from T₁plants are cultured and grown to maturity as T₂plants under the same conditions as used for culturing and growing the T₁plants. At least 10 independent transformation events are created from each construct for which T2 seeds are collected. The introduction of the gene is determined for each event by PCR performed on genomic DNA extracted from each event produced.
Transformation of tomato (Var M82) plants with putative cotton genes—Tomato (Solanum esculentum, var M82) transformation and cultivation of transgenic plants is effected according to: “Curtis I. S, Davey M. R, and Power J. B. 1995. “Leaf disk transformation”. Methods Mol. Biol. 44, 59-70 and Meissner R, Chague V, Zhu Q, Emmanuel E, Elkind Y, Levy A. A. 2000. “Technical advance: a high throughput system for transposon tagging and promoter trapping in tomato”. Plant J. 22, 265-74; with slight modifications.

Example 5

Evaluating Transgenic Arabidopsis Plant Growth Under Abiotic Stress as well as Under Favorable Conditions in Tissue Culture Assay

Assay I: plant growth under osmotic stress [poly (ethylene glycol) (PEG)] in tissue culture conditions—One of the consequences of drought is the induction of osmotic stress in the area surrounding the roots; therefore, in many scientific studies, PEG (e.g., 25% PEG8000) is used to simulate the osmotic stress conditions resembling the high osmolarity found during drought stress.
Surface sterilized seeds were sown in basal media [50% Murashige-Skoog medium (MS) supplemented with 0.8% plant agar as solidifying agent] in the presence of Kanamycin (for selecting only transgenic plants). After sowing, plates were transferred for 2-3 days for stratification at 4° C. and then grown at 25° C. under 12-hour light 12-hour dark daily cycles for 7 to 10 days. At this time point, seedlings randomly chosen were carefully transferred to plates containing 25% PEG: 0.5 MS media or Normal growth conditions (0.5 MS media). Each plate contained 5 seedlings of the same transgenic event, and 3-4 different plates (replicates) for each event. For each polynucleotide of the invention at least four independent transformation events were analyzed from each construct. Plants expressing the polynucleotides of the invention were compared to the average measurement of the control plants (empty vector or GUS reporter gene under the same promoter) used in the same experiment.
Digital imaging—A laboratory image acquisition system, which consists of a digital reflex camera (Canon EOS 300D) attached with a 55 mm focal length lens (Canon EF-S series), mounted on a reproduction device (Kaiser RS), which included 4 light units (4×150 Watts light bulb) and located in a darkroom, was used for capturing images of plantlets sawn in agar plates.
The image capturing process was repeated every 2-5 days starting at day 1 till day 10-15 (see for example the images in FIGS. 2A-B)
An image analysis system was used, which consists of a personal desktop computer (Intel P4 3.0 GHz processor) and a public domain program—ImageJ 1.39 (Java based image processing program which was developed at the U.S. National Institutes of Health and freely available on the internet at Hypertext Transfer Protocol://rsbweb (dot) nih (dot) gov/). Images were captured in resolution of 10 Mega Pixels (3888×2592 pixels) and stored in a low compression JPEG (Joint Photographic Experts Group standard) format. Next, analyzed data was saved to text files and processed using the JMP statistical analysis software (SAS institute).
Seedling analysis—Using the digital analysis seedling data was calculated, including leaf area, root coverage and root length.
The relative growth rate for the various seedling parameters was calculated according to the following formulas II, III and IV.
Relative growth rate of leaf area=(× rosette area/×t)*(1/rosette area t ₁) Formula II
Δ rosette area is the interval between the current rosette area (measured at t₂) and the rosette area measured at the previous day (Area t₁)
Δt is the time interval (t₂-t₁, in days) between the current analyzed image day (t₂) and the previous day (t₁).
Thus, the relative growth rate of leaf area is in units of 1/day.
Relative growth rate of root coverage=(Δ root coverage area/Δt)*(1/root coverage area t ₁) Formula III
Δ root coverage area is the interval between the current root coverage area (measured at t₂) and the root coverage area measured at the previous day (Area t₁)
Δt is the time interval (t₂-t₁, in days) between the current analyzed image day (t₂) and the previous day (t₁).
Thus, the relative growth rate of root coverage area is in units of 1/day.
Relative growth rate of root length=(Δ root length/Δt)*(1/root length t ₁) Formula IV
Δ root length is the interval between the current root length (measured at t₂) and the root length measured at the previous day (Area t₁)
Δt is the time interval (t₂-t₁, in days) between the current analyzed image day (t₂) and the previous day (t₁).
Thus, the relative growth rate of root length is in units of 1/day.
At the end of the experiment, plantlets were removed from the media and weighed for the determination of plant fresh weight. Plantlets were then dried for 24 hours at 60° C., and weighed again to measure plant dry weight for later statistical analysis. Growth rate was determined by comparing the leaf area coverage, root coverage and root length, between each couple of sequential photographs, and results were used to resolve the effect of the gene introduced on plant vigor, under osmotic stress, as well as under optimal conditions. Similarly, the effect of the gene introduced on biomass accumulation, under osmotic stress as well as under optimal conditions, was determined by comparing the plants' fresh and dry weight to that of control plants (containing an empty vector or the GUS reporter gene under the same promoter). From every construct created, 3-5 independent transformation events were examined in replicates.
Statistical analyses—To identify genes conferring significantly improved tolerance to abiotic stresses or enlarged root architecture, the results obtained from the transgenic plants were compared to those obtained from control plants. To identify outperforming genes and constructs, results from the independent transformation events tested were analyzed separately. To evaluate the effect of a gene event over a control the data was analyzed by Student's t-test and the p value was calculated. Results wer considered significant if p≦0.1. The JMP statistics software package was used (Version 5.2.1, SAS Institute Inc., Cary, N.C., USA).
Experimental Results—The polynucleotide sequences of the invention were assayed for a number of desired traits.
Tables 10-14 depict analyses of the above mentioned growth parameters of seedlings overexpres sing the polynucleotides of the invention under the regulation of the At6669 promoter (SEQ ID NO:2823) when grown under osmotic stress (25% PEG) conditions.

TABLE 10

MAB70 - 25% PEG

Event No.

7971.3

7972.1

7974.1

	Control	A	P	A	P	A	P

RGR of Roots Coverage between day 5 and 10	0.16	0.29	0.00	0.31	0.00	0.30	0.02
RGR of Roots Length between day 1 and 5	0.07	0.12	0.05	0.13	0.03

Table 10: Provided are the growth and biomass parameters of transgenic or control plants as measured in Tissue Calture growth under 25% PEG.
A = average;
P = p value;
RGR = Relative Growth Rate.
The indicated days refer to days from planting.

TABLE 11

MAB76 - 25% PEG

Event No.

7635.4

7635.1

	Control	A	P	A	P

RGR of Roots Coverage	0.16	0.22	0.02	0.21	0.09
between day 5 and 10

Table 11: Provided are the growth and biomass parameters of transgenic or control plants as measured in Tissue Calture growth under 25% PEG. A = average; P = p value; RGR = Relative Growth Rate. The indicated days refer to days from planting.

TABLE 12

MAB79 - 25% PEG

Event No.

7324.1

7961.1

	Control	A	P	A	P

Dry Weight [gr]	0.01	0.01	0.06
Fresh Wight [gr]	0.08			0.13	0.07
Leaf Area on day 5	0.15			0.19	0.05
RGR of Roots Coverage	0.16			0.32	0.05
between day 5 and 10
RGR of Roots Length	0.07			0.11	0.02
between day 1 and 5

Table 12: Provided are the growth and biomass parameters of transgenic or control plants as measured in Tissue Calture growth under 25% PEG. A = average; P = p value; RGR = Relative Growth Rate. The indicated days to refer to days from planting.

TABLE 13

MAB56 - 25% PEG

Event No.

6802.10

	Control	A	P

Roots Coverage on day 7	2.46	3.38	0.09
Roots Length on day 7	2.81	3.43	0.07

Table 13: Provided are the growth and biomass parameters of transgenic or control plants as measured in Tissue Calture growth under 25% PEG. A = average; P = p value; RGR = Relative Growth Rate. The indicated days refer to days from planting.

TABLE 14

MAB58 - 25% PEG

Event No.

6783.30

	Control	A	P

Leaf Area on day 7	0.31	0.41	0.03
Leaf Area on day 14	0.80	1.09	0.02
Fresh Weight	0.18	0.31	0.00
Dry Weight [gr]	0.01	0.013	0.01

Table 14: Provided are the growth and biomass parameters of transgenic or control plants as measured in Tissue Calture growth under 25% PEG. A = average; P = p value; RGR = Relative Growth Rate. The indicated days refer to days from planting.

Tables 15-29 depict analyses of the above mentioned growth parameters of seedlings overexpres sing the polynucleotides of the invention under the regulation of the At6669 promoter in Normal Growth conditions (0.5 MS medium).

TABLE 15

MAB70 - Normal Growth Conditions

Event No.

7971.3

7972.1

7974.1

7974.3

	Control	A	P	A	P	A	P	A	P

Dry Weight [gr]	0.01	0.01	0.03	0.01	0.05	0.02	0.04	0.01	0.07
Fresh Wight [gr]	0.14					0.24	0.04	0.22	0.10
Leaf Area on day 10	0.46	0.59	0.00
Leaf Area on day 5	0.21					0.30	0.10
RGR of Roots Coverage between	0.18	0.42	0.00	0.37	0.00	0.32	0.01
day 5 and 10
RGR of Roots Length between day	0.06	0.15	0.01	0.16	0.00	0.15	0.00
1 and 5
RGR of Roots Length between day	0.22			0.32	0.09
5 and 10

Table 15: Provided are the growth and biomass parameters of transgenic or control plants as measured in Tissue Calture growth under normal growth conditions.
A = average;
P = p value;
RGR = Relative Growth Rate.
The indicated days refer to days from planting.

TABLE 16

MAB71 - Normal Growth Conditions

Event No.

7331.5

7332.2

7333.5

	Control	A	P	A	P	A	P

Dry Weight [gr]	0.01	0.01	0.05
Fresh Wight [gr]	0.14	0.20	0.08
RGR of Roots Coverage between day 5 and 10	0.18			0.28	0.04	0.26	0.07
RGR of Roots Length between day 1 and 5	0.06			0.11	0.02	0.11	0.01

Table 16: Provided are the growth and biomass parameters of transgenic or control plants as measured in Tissue Calture growth under normal growth conditions.
A = average;
P = p value;
RGR = Relative Growth Rate.
The indicated days refer to days from planting.

TABLE 17

MAB74 - Normal Growth Conditions

Event No.

7981.1

7982.4

7983.9

	Control	A	P	A	P	A	P

Dry Weight [gr]	0.01	0.01	0.09	0.02	0.05	0.01	0.06
Fresh Wight [gr]	0.14					0.21	0.01
Leaf Area on day 10	0.46	0.67	0.01
Leaf Area on day 5	0.21	0.29	0.04	0.29	0.01	0.27	0.00
RGR of Roots Coverage between day 5 and 10	0.18			0.31	0.07
RGR of Roots Coverage between day 1 and 5	0.73			1.70	0.07
RGR of Roots Length between day 1 and 5	0.06			0.12	0.09	0.14	0.03
RGR of Roots Length between day 5 and 10	0.22	0.45	0.05	0.58	0.00

Table 17: Provided are the growth and biomass parameters of transgenic or control plants as measured in Tissue Calture growth under normal growth conditions.
A = average;
P = p value;
RGR = Relative Growth Rate.
The indicated days refer to days from planting.

TABLE 18

MAB76 - Normal Growth Conditions

Event No.

7633.3

7635.4

7635.1

	Control	A	P	A	P	A	P

RGR Leaf Area	0.24	0.34	0.04	0.33	0.07
between day 5
and 10
RGR of Roots	0.18					0.38	0.08
Coverage between
day 5 and 10
RGR of Roots	0.06					0.13	0.02
Length between
day 1 and 5

Table 18: Provided are the growth and biomass parameters of transgenic or control plants as measured in Tissue Calture growth under normal growth conditions.
A = average;
P = p value;
RGR = Relative Growth Rate.
The indicated days refer to days from planting.

TABLE 19

MAB77 - Normal Growth Conditions

Event No.

7931.11

8211.2

8212.2

	Control	A	P	A	P	A	P

Dry Weight [gr]	0.01	0.01	0.08			0.01	0.10
Roots Coverage on day 10	2.77					4.33	0.07
RGR Leaf Area between day 5 and 10	0.24	0.38	0.10	0.35	0.04
RGR of Roots Coverage between day 5 and 10	0.18	0.27	0.09	0.29	0.09
RGR of Roots Length between day 1 and 5	0.06			0.10	0.03

Table 19: Provided are the growth and biomass parameters of transgenic or control plants as measured in Tissue Calture growth under normal growth conditions.
A = average;
P = p value;
RGR = Relative Growth Rate.
The indicated days refer to days from planting.

TABLE 20

MAB79 - Normal Growth Conditions

Event No.

7323.3

7324.1

7961.1

7962.2

	Control	A	P	A	P	A	P	A	P

Dry Weight [gr]	0.01	0.02	0.03	0.02	0.08	0.01	0.00
Fresh Wight [gr]	0.14	0.34	0.04	0.31	0.09
Leaf Area on day 10	0.46	0.92	0.01	0.90	0.01	0.81	0.00
Leaf Area on day 5	0.21					0.29	0.02	0.28	0.00
Roots Coverage on day 10	2.77	5.31	0.02	4.59	0.07	3.81	0.00	3.71	0.01
RGR Leaf Area between day 5 and	0.24					0.36	0.02
10
RGR Leaf Area between day 1 and 5	0.82					1.34	0.00
RGR of Roots Coverage between	0.18					0.45	0.06	0.39	0.02
day 5 and 10
RGR of Roots Length between day	0.06	0.12	0.02	0.17	0.06			0.14	0.00
1 and 5

Table 20: Provided are the growth and biomass parameters of transgenic or control plants as measured in Tissue Calture growth under normal growth conditions.
A = average;
P = p value;
RGR = Relative Growth Rate.
The indicated days refer to days from planting.

TABLE 21

MAB115 - Normal Growth Conditions

Event No.

8561.2..

8564.1..

8564.2..

8565.1..

	Control	A	P	A	P	A	P	A	P

Dry Weight [gr]	0.005	0.006	0.00					0.009	0.00
Leaf Area on day 10	0.24							0.27	0.00
Leaf Area on day 5	0.35							0.38	0.01
Roots Coverage on day 10	1.67							2.13	0.02
Roots Coverage on day 5	3.38							4.80	0.03
Roots Length on day 10	2.39							2.74	0.00
Roots Length on day 5	3.46							4.22	0.02
RGR Leaf Area between day 5	0.37	0.43	0.00					0.48	0.00
and 10
RGR Leaf Area between day 1	0.16	0.19	0.00
and 5
RGR of Roots Coverage	1.89	3.21	0.00	2.24	0.02	2.23	0.02	3.47	0.01
between day 5 and 10
RGR of Roots Coverage	0.33	0.35	0.00	0.41	0.00	0.43	0.00	0.44	0.00
between day 1 and 5
RGR of Roots Length between	0.37	0.56	0.02			0.53	0.06	0.68	0.00
day 1 and 5
RGR of Roots Length between	0.15			0.18	0.00	0.17	0.00	0.18	0.00
day 5 and 10

Table 21: Provided are the growth and biomass parameters of transgenic or control plants as measured in Tissue Calture growth under normal growth conditions.
A = average;
P = p value;
RGR = Relative Growth Rate.
The indicated days refer to days from planting.

TABLE 22

MAB54 - Normal Growth Conditions

Event No.

8181.2..

8182.2..

8184.3..

8185.3..

	Control	A	P	A	P	A	P	A	P

Dry Weight [gr]	0.005	0.009	0.00	0.008	0.00	0.008	0.00	0.007	0.00
Roots Coverage on day 10	1.67	1.80	0.04
Roots Coverage on day 5	3.38	4.27	0.02	3.40	0.00
Roots Length on day 10	2.39	2.52	0.01
Roots Length on day 5	3.46	4.11	0.02	3.50	0.00
RGR Leaf Area between day 5	0.37	0.45	0.00
and 10
RGR Leaf Area between day 1	0.16	0.17	0.04					0.19	0.00
and 5
RGR of Roots Coverage	1.89	3.59	0.00	3.60	0.01	2.28	0.01	2.20	0.01
between day 5 and 10
RGR of Roots Coverage	0.33	0.52	0.00	0.55	0.00	0.39	0.00	0.36	0.00
between day 1 and 5
RGR of Roots Length between	0.37	0.70	0.00	0.73	0.00	0.48	0.08	0.51	0.02
day 1 and 5
RGR of Roots Length between	0.15	0.21	0.01	0.22	0.01	0.17	0.00	0.17	0.00
day 5 and 10

Table 22: Provided are the growth and biomass parameters of transgenic or control plants as measured in Tissue Calture growth under normal growth conditions.
A = average;
P = p value;
RGR = Relative Growth Rate.
The indicated days refer to days from planting.

TABLE 23

MAB55 - Normal Growth Conditions

Event No.

6802.1

6802.11

6802.8

6805.3

	A	P	A	P	A	P	A	P

Roots Coverage on day 7	3.67	5.93	0.04
Roots Coverage on day 14	7.40	13.26	0.04
Roots Length on day 7	3.99	5.32	0.01
Roots Length on day 14	6.14	7.80	0.04					7.65	0.04
RGR of Roots Coverage between day	0.53	1.30	0.00	1.11	0.02	0.93	0.02
1 and 7
RGR of Roots Length between day 1	0.29	0.48	0.00	0.45	0.03	0.43	0.02
and 7
Fresh Weight	0.19					0.10	0.00	0.12	0.01
Dry Weight [gr]	0.01					0.01	0.00	0.01	0.00

Table 23: Provided are the growth and biomass parameters of transgenic or control plants as measured in Tissue Calture growth under normal growth conditions.
A = average;
P = p value;
RGR = Relative Growth Rate.
The indicated days refer to days from planting.

TABLE 24

MAB56 - Normal Growth Conditions

Event No.

6691.2

6691.3

6693.2

6693.5

	A	P	A	P	A	P	A	P

Roots Coverage on day 7	3.67	5.81	0.00					5.67	0.01
Roots Length on day 7	3.99	6.21	0.00					5.39	0.00
Roots Length on day 14	6.14	8.32	0.01					8.01	0.02
RGR of Roots Length between day 7	0.09	0.05	0.01	0.05	0.06	0.06	0.08
and 14
Fresh Weight	0.19	0.14	0.03	0.14	0.03
Dry Weight [gr]	0.01	0.01	0.02	0.01	0.02	0.00	0.00

Table 24: Provided are the growth and biomass parameters of transgenic or control plants as measured in Tissue Calture growth under normal growth conditions.
A = average;
P = p value;
RGR = Relative Growth Rate.
The indicated days refer to days from planting.

TABLE 25

MAB57 - Normal Growth Conditions

Event No.

6912.14

6912.2

6912.6

6914.1

	A	P	A	P	A	P	A	P

Roots Coverage on day 7	3.67	6.40	0.00	6.71	0.00
Roots Coverage on day 14	7.40	17.33	0.00	15.27	0.05	12.30	0.02
Roots Length on day 7	3.99	5.54	0.00	6.12	0.00	6.34	0.00
Roots Length on day 14	6.14	8.76	0.00	8.48	0.01	7.97	0.02	7.85	0.04
RGR of Roots Length between day	0.29	0.39	0.06
1 and 7
RGR of Roots Length between day	0.09					0.04	0.09
7 and 14
Fresh Weight	0.19	0.24	0.09			0.11	0.01	0.11	0.01
Dry Weight [gr]	0.01					0.01	0.01	0.01	0.01

Table 25: Provided are the growth and biomass parameters of transgenic or control plants as measured in Tissue Calture growth under normal growth conditions.
A = average;
P = p value;
RGR = Relative Growth Rate.
The indicated days refer to days from planting.

TABLE 26

MAB58 - Normal Growth Conditions

Event No.

6783.1

6783.2

6783.3

	A	P	A	P	A	P

Roots Coverage on	3.67					6.18	0.00
day 7
Roots Coverage on	7.40			12.65	0.02	12.00	0.09
day 14
Roots Length on day 7	3.99					5.20	0.02
Roots Length on	6.14	7.38	0.09	7.51	0.07	7.59	0.08
day 14
RGR of Roots Length	0.29	0.35	0.07
between day 1 and 7
Fresh Weight	0.19	0.13	0.03
Dry Weight [gr]	0.01	0.01	0.00

Table 26: Provided are the growth and biomass parameters of transgenic or control plants as measured in Tissue Calture growth under normal growth conditions.
A = average;
P = p value;
RGR = Relative Growth Rate.
The indicated days refer to days from planting.

TABLE 27

MAB59 - Normal Growth Condition

Event No.

6791.4

6793.4

6794.4

	A	P	A	P	A	P

Roots Coverage on	3.67			5.61	0.04	5.23	0.09
day 7
Roots Coverage on	7.40			9.65	0.09	10.28	0.09
day 14
Roots Length on day 7	3.99			5.47	0.08	4.95	0.09
Roots Length on	6.14					7.70	0.07
day 14
RGR of Roots	0.53					0.80	0.09
Coverage between
day 1 and 7
RGR of Roots Length	0.09			0.05	0.10
between day 7 and 14
Fresh Weight	0.19	0.09	0.00
Dry Weight [gr]	0.01	0.004	0.00	0.005	0.02

Table 27: Provided are the growth and biomass parameters of transgenic or control plants as measured in Tissue Calture growth under normal growth conditions.
A = average;
P = p value;
RGR = Relative Growth Rate.
The indicated days refer to days from planting.

TABLE 28

MAB69 - Normal Growth Conditions

Event No.

6651.1

6651.12

6651.13

6651.8

	A	P	A	P	A	P	A	P

Roots Length on day 7	3.99	4.97	0.10
Roots Length on day 14	6.14	8.01	0.02
RGR of Roots Length between day	0.29							0.35	0.09
1 and 7
Fresh Weight	0.19					0.12	0.02	0.12	0.01
Dry Weight [gr]	0.01			0.007	0.09	0.005	0.00	0.006	0.00

Table 28: Provided are the growth and biomass parameters of transgenic or control plants as measured in Tissue Calture growth under normal growth conditions.
A = average;
P = p value;
RGR = Relative Growth Rate.
The indicated days refer to days from planting.

TABLE 29

MAB72 - Normal Growth Conditions

Event No.

8552.1..

8552.4..

8553.2..

	Control	A	P	A	P	A	P

Dry Weight [gr]	0.005	0.008	0.00	0.008	0.00	0.007	0.00
Leaf Area on day 10	0.24	0.24	0.01
Roots Coverage on day 10	1.67	1.84	0.01	1.93	0.02	1.89	0.03
Roots Coverage on day 5	3.38	3.60	0.04	3.90	0.06	4.50	0.00
Roots Length on day 10	2.39	2.48	0.01
Roots Length on day 5	3.46	3.70	0.04	3.65	0.04	3.84	0.00
RGR Leaf Area between day 5 and 10	0.37	0.47	0.00	0.39	0.00
RGR Leaf Area between day 1 and 5	0.16			0.20	0.09
RGR of Roots Coverage between day 5 and	1.89	1.93	0.03	2.29	0.06	3.04	0.01
10
RGR of Roots Coverage between day 1 and 5	0.33			0.35	0.00	0.52	0.00
RGR of Roots Length between day 1 and 5	0.37					0.55	0.01
RGR of Roots Length between day 5 and 10	0.15	0.16	0.00	0.18	0.00	0.22	0.01

Table 29: Provided are the growth and biomass parameters of transgenic or control plants as measured in Tissue Calture growth under normal growth conditions.
A = average;
P = p value;
RGR = Relative Growth Rate.
The indicated days refer to days from planting.

Example 6

Evaluating Transgenic Arabidopsis Plant Growth Under Abiotic Stress as well as Favorable Conditions in Greenhouse Assay

ABS tolerance: Yield and plant growth rate at high salinity concentration under greenhouse conditions—This assay followed the rosette area growth of plants grown in the greenhouse as well as seed yield at high salinity irrigation. Seeds were sown in agar media supplemented only with a selection agent (Kanamycin) and Hoagland solution under nursery conditions. The T₂transgenic seedlings were then transplanted to 1.7 trays filled with peat and perlite. The trays were irrigated with tap water (provided from the pots' bottom). Half of the plants were irrigated with a salt solution (40-80 mM NaCl and 5 mM CaCl₂) so as to induce salinity stress (stress conditions). The other half of the plants was irrigated with tap water (normal conditions). All plants were grown in the greenhouse until mature seeds, then harvested (the above ground tissue) and weighted (immediately or following drying in oven at 50° C. for 24 hours). High salinity conditions were achieved by irrigating with a solution containing 40-80 mM NaCl (“ABS” growth conditions) and compared to regular growth conditions.
Each construct was validated at its T2 generation. Transgenic plants transformed with a construct including the uidA reporter gene (GUS) under the AT6669 promoter or with an empty vector including the AT6669 promoter were used as control.
The plants were analyzed for their overall size, growth rate, flowering, seed yield, weight of 1,000 seeds, dry matter and harvest index (HI—seed yield/dry matter). Transgenic plants performance was compared to control plants grown in parallel under the same conditions. Mock—transgenic plants expressing the uidA reporter gene (GUS-Intron) or with no gene at all, under the same promoter were used as control.
The experiment was planned in nested randomized plot distribution. For each gene of the invention three to five independent transformation events were analyzed from each construct.
Digital imaging—A laboratory image acquisition system, which consists of a digital reflex camera (Canon EOS 300D) attached with a 55 mm focal length lens (Canon EF-S series), mounted on a reproduction device (Kaiser RS), which included 4 light units (4×150 Watts light bulb) was used for capturing images of plant samples.
The image capturing process was repeated every 2 days starting from day 1 after transplanting till day 16. Same camera, placed in a custom made iron mount, was used for capturing images of larger plants sawn in white tubs in an environmental controlled greenhouse. The tubs were square shape include 1.7 liter trays. During the capture process, the tubs were placed beneath the iron mount, while avoiding direct sun light and casting of shadows.
An image analysis system was used, which consists of a personal desktop computer (Intel P4 3.0 GHz processor) and a public domain program—ImageJ 1.39 (Java based image processing program which was developed at the U.S. National Institutes of Health and freely available on the internet at Hypertext Transfer Protocol://rsbweb (dot) nih (dot) gov/). Images were captured in resolution of 10 Mega Pixels (3888×2592 pixels) and stored in a low compression JPEG (Joint Photographic Experts Group standard) format. Next, analyzed data was saved to text files and processed using the JMP statistical analysis software (SAS institute).
Leaf analysis—Using the digital analysis leaves data was calculated, including leaf number, area, perimeter, length and width.
Vegetative growth rate: the relative growth rate (RGR) of leaf number and rosette area were calculated formulas V and VI, respectively.
Relative growth rate of leaf number=(Δ leaf number/Δt)*(1/leaf number t ₁) Formula V
Δ leaf number is the interval between the current leaf number (measured at t₂) and the leaf number measured at the previous day (Area t₁)
Δt is the time interval (t₂-t₁, in days) between the current analyzed image day (t₂) and the previous day (t₁).
Thus, the relative growth rate of leaf number is in units of 1/day.
Relative growth rate of rosette area=(Δ rosette area/Δt)*(1/rosette area t ₁) Formula VI
Δ rosette area is the interval between the current rosette area (measured at t₂) and the rosette area measured at the previous day (Area t₁)
Δt is the time interval (t₂-t₁, in days) between the current analyzed image day (t₂) and the previous day (t₁).
Thus, the relative growth rate of rosette area is in units of 1/day.
Seeds average weight—At the end of the experiment all seeds were collected. The seeds were scattered on a glass tray and a picture was taken. Using the digital analysis, the number of seeds in each sample was calculated.
Dry weight and seed yield—On about day 80 from sowing, the plants were harvested and left to dry at 30° C. in a drying chamber. The biomass and seed weight of each plot were measured and divided by the number of plants in each plot. Dry weight=total weight of the vegetative portion above ground (excluding roots) after drying at 30° C. in a drying chamber; Seed yield per plant=total seed weight per plant (gr). 1000 seed weight (the weight of 1000 seeds) (gr.).
Harvest Index (HI)—The harvest index was calculated using Formula VII.
Harvest Index=Average seed yield per plant/Average dry weight Formula VII
Statistical analyses—To identify genes conferring significantly improved tolerance to abiotic stresses, the results obtained from the transgenic plants were compared to those obtained from control plants. To identify outperforming genes and constructs, results from the independent transformation events tested were analyzed separately. Data was analyzed using Student's t-test and results were considered significant if the p value was less than 0.1. The JMP statistics software package is used (Version 5.2.1, SAS Institute Inc., Cary, N.C., USA).
Experimental Results
Tables 30-45 depict analyses of plant parameters as describe above overexpressing the polynucleotides of the invention under the regulation of the At6669 promoter under salinity irrigation conditions [NaCl 40-80 mM; NaCl Electrical conductivity (E.C.) of 7-10].

TABLE 30

MAB115 - Salt irrigation (40-80 mM NaCl)

Event No.

8564.1

8565.1

	Control	A	P	A	P

Rosette Diameter on day 3*	1.70	1.80	0.09	1.75	0.04
Rosette Diameter on day 5	2.36
Rosette Diameter on day 8	3.77			4.00	0.09
Rosette Area on day 3	0.90
Rosette Area on day 5	1.56			1.70	0.09
Rosette Area on day 8	4.06			4.38	0.06
Plot Coverage on day 5	12.30			13.61	0.06
Plot Coverage on day 8	31.90			35.07	0.02
Leaf Number on day 3	5.08			5.94	0.00
Leaf Number on day 5	6.86			7.38	0.05
RGR of Leaf Number between	0.18
day 3 and 5
RGR of Leaf Number between	0.09
day 5 and 8
RGR of Rosette Area between	0.53	0.62	0.01
day 5 and 8
Biomass DW [gr]	3.24
Harvest Index	0.11	0.15	0.07	0.16	0.03

Table 30: Provided are the growth, biomass and yield parameters of transgenic or control plants as measured in Green House under salinity irrigation. A = average; P = p value; RGR = Relative Growth Rate. The indicated days refer to days from planting.

TABLE 31

MAB54 - Salt irrigation (40-80 mM NaCl)

Event No.

8181.2

8182.2

8183.2

8185.4

	Control	A	P	A	P	A	P	A	P

Rosette Diameter on day 3*	1.70	1.95	0.04
Rosette Diameter on day 5	2.36			2.70	0.02	2.64	0.00
Rosette Diameter on day 8	3.77			4.00	0.08
Rosette Area on day 3	0.90					1.19	0.04
Plot Coverage on day 3	7.04					9.52	0.04	7.49	0.08
Leaf Number on day 3	5.08			6.19	0.06
Leaf Number on day 8	8.66	9.31	0.00	9.38	0.00
RGR of Leaf Number between day	0.18
3 and 5
RGR of Rosette Area between day	0.45			0.52	0.01
1 and 3
1000 Seeds weight [gr]	0.02					0.02	0.00	0.02	0.00
Yield [gr]/Plant	0.04	0.06	0.05
Harvest Index	0.11	0.17	0.01

Table 31: Provided are the growth, biomass and yield parameters of transgenic or control plants as measured in Green House under salinity irrigation.
A = average;
P = p value;
RGR = Relative Growth Rate.
The indicated days refer to days from planting.

TABLE 32

MAB55 - Salt irrigation (40-80 mM NaCl)

Event No.

6802.10

6805.3

6805.4

	Control	A	P	A	P	A	P

Rosette Diameter on day 5	2.36	2.81	0.03
Rosette Diameter on day 8	3.77	4.29	0.05
Rosette Area on day 3	0.90	1.35	0.01
Rosette Area on day 5	1.56			1.70	0.03
Rosette Area on day 8	4.06	5.91	0.05
Plot Coverage on day 3	7.04	10.76	0.01
Plot Coverage on day 5	12.30			13.60	0.02
Plot Coverage on day 8	31.90	47.28	0.06
Leaf Number on day 3	5.08	6.25	0.00
Leaf Number on day 5	6.86	7.94	0.03
Leaf Number on day 8	8.66	9.50	0.01
RGR of Rosette Area between day 1 and 3	0.45	0.51	0.05
Yield [gr]/Plant	0.04					0.06	0.03
Harvest Index	0.11			0.15	0.05

Table 32: Provided are the growth, biomass and yield parameters of transgenic or control plants as measured in Green House under salinity irrigation.
A = average;
P = p value;
RGR = Relative Growth Rate.
The indicated days refer to days from planting.

TABLE 33

MAB56 - Salt irrigation (40-80 mM NaCl)

Event No.

6691.2

6691.3

6693.2

6695.6

	Control	A	P	A	P	A	P	A	P

Rosette Diameter on day 3	1.70					1.89	0.05	1.97	0.07
Rosette Diameter on day 5	2.36					2.55	0.09	2.58	0.01
Rosette Area on day 3	0.90			1.06	0.00	1.15	0.02	1.23	0.08
Rosette Area on day 5	1.56					1.94	0.02	1.94	0.03
Rosette Area on day 8	4.06	4.63	0.02
Plot Coverage on day 3	7.04			8.45	0.00	9.21	0.01	9.82	0.07
Plot Coverage on day 5	12.30					15.49	0.01	15.53	0.02
Plot Coverage on day 8	31.90	37.03	0.01					34.82	0.05
Leaf Number on day 3	5.08			5.50	0.00	5.81	0.00	6.00	0.02
Leaf Number on day 5	6.86					7.38	0.01	7.50	0.02
1000 Seeds weight [gr]	0.02			0.02	0.08
Harvest Index	0.11			0.18	0.01

Table 33: Provided are the growth, biomass and yield parameters of transgenic or control plants as measured in Green House under salinity irrigation.
A = average;
P = p value;
RGR = Relative Growth Rate.
The indicated days refer to days from planting.

TABLE 34

MAB57 - Salt irrigation (40-80 mM NaCl)

Event No.

6912.1

6912.13

6914.5

	Control	A	P	A	P	A	P

Leaf Number on day 3	5.08					5.69	0.00
Leaf Number on day 5	6.86			8.13	0.06
Leaf Number on day 8	8.66					9.56	0.06
RGR of Leaf Number between day 5 and 8	0.09	0.14	0.01
RGR of Rosette Area between day 5 and 8	0.53	0.62	0.02			0.58	0.10
1000 Seeds weight [gr]	0.02					0.02	0.00
Yield [gr]/Plant	0.04	0.06	0.01	0.06	0.03	0.06	0.01
Harvest Index	0.11			0.19	0.06	0.23	0.00

Table 34: Provided are the growth, biomass and yield parameters of transgenic or control plants as measured in Green House under salinity irrigation.
A = average;
P = p value;
RGR = Relative Growth Rate.
The indicated days refer to days from planting.

TABLE 35

MAB58 - Salt irrigation (40-80 mM NaCl)

Event No.

6783.3

7522.10

7522.3

7523.6

	Control	A	P	A	P	A	P	A	P

Rosette Diameter on day 3	1.70							2.11	0.00
Rosette Diameter on day 5	2.36	2.57	0.00
Rosette Diameter on day 8	3.77					4.06	0.07	4.54	0.00
Rosette Area on day 3	0.90							1.36	0.00
Rosette Area on day 5	1.56							2.29	0.00
Rosette Area on day 8	4.06							5.98	0.00
Plot Coverage on day 3	7.04							10.90	0.00
Plot Coverage on day 5	12.30							18.32	0.00
Plot Coverage on day 8	31.90							47.88	0.00
Leaf Number on day 3	5.08			5.63	0.06			6.06	0.00
Leaf Number on day 8	8.66	9.25	0.04					9.81	0.04
RGR of Leaf Number between day	0.09	0.12	0.03
5 and 8
1000 Seeds weight [gr]	0.02	0.02	0.08
Yield [gr]/Plant	0.04	0.06	0.09

Table 35: Provided are the growth, biomass and yield parameters of transgenic or control plants as measured in Green House under salinity irrigation.
A = average;
P = p value;
RGR = Relative Growth Rate.
The indicated days refer to days from planting.

TABLE 36

MAB59 - Salt irrigation (40-80 mM NaCl)

Event No.

6791.6

6794.4

6794.5

	Control	A	P	A	P	A	P

Rosette Diameter on	1.70					2.38	0.05
day 3
Rosette Diameter on	2.36			2.73	0.06
day 5
Rosette Area on	0.90			1.27	0.06	1.65	0.03
day 3
Plot Coverage on	7.04			10.15	0.06	13.19	0.03
day 3
Leaf Number on	5.08	6.44	0.05	6.00	0.02	6.75	0.01
day 3
Leaf Number on	6.86	8.38	0.00	7.69	0.05	8.00	0.07
day 5
Leaf Number on	8.66					9.38	0.02
day 8
Yield [gr]/Plant	0.04	0.06	0.06
Harvest Index	0.11	0.16	0.04

Table 36: Provided are the growth, biomass and yield parameters of transgenic or control plants as measured in Green House under salinity irrigation.
A = average;
P = p value;
RGR = Relative Growth Rate.
The indicated days refer to days from planting.

TABLE 37

MAB69 - Salt irrigation (40-80 mM NaCl)

Event No.

6651.11

6651.12

6651.2

8342.1

	Control	A	P	A	P	A	P	A	P

Rosette Diameter on day 3	1.70			1.92	0.01
Rosette Area on day 3	0.90			1.09	0.00
Rosette Area on day 8	4.06			5.05	0.04
Plot Coverage on day 3	7.04			8.74	0.00
Plot Coverage on day 8	31.90			40.41	0.04
Leaf Number on day 3	5.08			5.69	0.00
Leaf Number on day 8	8.66							8.94	0.08
RGR of Rosette Area between day	0.45	0.49	0.02	0.51	0.04
1 and 3
1000 Seeds weight [gr]	0.02	0.02	0.00			0.02	0.01
Yield [gr]/Plant	0.04	0.05	0.09	0.06	0.02			0.07	0.01
Harvest Index	0.11			0.17	0.09	0.22	0.01

Table 38: Provided are the growth, biomass and yield parameters of transgenic or control plants as measured in Green House under salinity irrigation.
A = average;
P = p value;
RGR = Relative Growth Rate.
The indicated days refer to days from planting.

TABLE 39

MAB70 - Salt irrigation (40-80 mM NaCl)

Event No.

7971.3

7972.1

7972.3

	Control	A	P	A	P	A	P

Rosette Diameter on day 3	1.70					1.94	0.00
Rosette Diameter on day 5	2.36					2.62	0.04
Rosette Diameter on day 8	3.77			4.40	0.00
Rosette Area on day 3	0.90			1.20	0.07	1.12	0.01
Rosette Area on day 5	1.56			2.06	0.05	1.87	0.00
Rosette Area on day 8	4.06			5.86	0.06
Plot Coverage on day 3	7.04			9.61	0.06	9.00	0.01
Plot Coverage on day 5	12.30			16.46	0.04	14.93	0.00
Plot Coverage on day 8	31.90			46.87	0.06
Leaf Number on day 3	5.08	5.94	0.09
Leaf Number on day 8	8.66	9.31	0.00			9.75	0.09
RGR of Rosette Area between day 5 and 8	0.53			0.62	0.01
Yield [gr]/Plant	0.04	0.08	0.00	0.07	0.01
Harvest Index	0.11	0.25	0.00

Table 39: Provided are the growth, biomass and yield parameters of transgenic or control plants as measured in Green House under salinity irrigation.
A = average;
P = p value;
RGR = Relative Growth Rate.
The indicated days refer to days from planting.

TABLE 40

MAB71 - Salt irrigation (40-80 mM NaCl)

Event No.

7331.4

7331.5

7333.5

7334.5

	Control	A	P	A	P	A	P	A	P

Rosette Diameter on day 5	2.36	3.03	0.00			2.52	0.02
Rosette Diameter on day 8	3.77	5.01	0.05	4.33	0.01
Plot Coverage on day 5	12.30	19.90	0.09			14.25	0.08
Leaf Number on day 3	5.08	6.44	0.05			5.47	0.06
Leaf Number on day 5	6.86	8.13	0.00	7.56	0.00
Leaf Number on day 8	8.66	10.38	0.05
RGR of Leaf Number between	0.09							0.12	0.01
day 5 and 8
RGR of Rosette Area between	0.53	0.61	0.03					0.61	0.04
day 5 and 8
Yield [gr]/Plant	0.04							0.05	0.10
Harvest Index	0.11	0.21	0.06

Table 40: Provided are the growth, biomass and yield parameters of transgenic or control plants as measured in Green House under salinity irrigation.
A = average;
P = p value;
RGR = Relative Growth Rate.
The indicated days refer to days from planting.

TABLE 41

MAB72 - Salt irrigation (40-80 mM NaCl)

Event No.

8552.1

8553.2

8553.3

8555.3

	Control	A	P	A	P	A	P	A	P

Rosette Diameter on day 3	1.70			1.90	0.05			1.83	0.00
Rosette Diameter on day 5	2.36			2.67	0.00			2.55	0.01
Rosette Diameter on day 8	3.77			4.06	0.08	4.15	0.10
Rosette Area on day 3	0.90			1.20	0.00	1.02	0.00	1.08	0.02
Rosette Area on day 5	1.56	1.88	0.07	2.00	0.00			1.87	0.00
Rosette Area on day 8	4.06	4.68	0.00	5.05	0.00			4.82	0.00
Plot Coverage on day 3	7.04			9.64	0.00	8.13	0.00	8.67	0.02
Plot Coverage on day 5	12.30	15.05	0.05	16.04	0.00			14.98	0.00
Plot Coverage on day 8	31.90	37.48	0.00	40.39	0.00			38.57	0.00
Leaf Number on day 3	5.08	5.81	0.00	5.88	0.00	5.56	0.00	5.50	0.00
Leaf Number on day 8	8.66					9.38	0.02
RGR of Leaf Number between	0.12	0.17	0.01
day 1 and 3

Table 41: Provided are the growth, biomass and yield parameters of transgenic or control plants as measured in Green House under salinity irrigation.
A = average;
P = p value;
RGR = Relative Growth Rate.
The indicated days refer to days from planting.

TABLE 42

MAB74 - Salt irrigation (40-80 mM NaCl)

Event No.

7982.1

7983.6

7983.9

	Control	A	P	A	P	A	P

Rosette Diameter on day 3	1.70			2.06	0.10	1.94	0.00
Rosette Diameter on day 5	2.36					2.62	0.06
Rosette Diameter on day 8	3.77					4.05	0.02
Rosette Area on day 3	0.90			1.14	0.02
Rosette Area on day 5	1.56			1.83	0.05	1.85	0.10
Rosette Area on day 8	4.06					4.46	0.03
Plot Coverage on day 3	7.04			9.13	0.02
Plot Coverage on day 5	12.30			14.62	0.03	14.79	0.08
Plot Coverage on day 8	31.90					35.66	0.01
Leaf Number on day 3	5.08			5.75	0.04	5.31	0.07
Leaf Number on day 5	6.86					7.25	0.04
RGR of Rosette Area between day 3 and 5	0.37	0.42	0.07
1000 Seeds weight [gr]	0.02	0.02	0.02			0.02	0.05
Yield [gr]/Plant	0.04	0.06	0.05
Harvest Index	0.11	0.20	0.06

Table 42: Provided are the growth, biomass and yield parameters of transgenic or control plants as measured in Green House under salinity irrigation.
A = average;
P = p value;
RGR = Relative Growth Rate.
The indicated days refer to days from planting.

TABLE 43

MAB76 - Salt irrigation (40-80 mM NaCl)

Event No.

7633.1

7633.2

	Control	A	P	A	P

Leaf Number on day 3	5.08			5.44	0.02
Leaf Number on day 8	8.66	9.13	0.07

Table 43: Provided are the growth, biomass and yield parameters of transgenic or control plants as measured in Green House under salinity irrigation. A = average; P = p value; RGR = Relative Growth Rate. The indicated days refer to days from planting.

TABLE 44

MAB77 - Salt irrigation (40-80 mM NaCl)

Event No.

7931.11

8212.2

	Control	A	P	A	P

Leaf Number on day 8	8.66			9.75	0.09
RGR of Rosette Area	0.45	0.52	0.10
between day 1 and 3
Harvest Index	0.11			0.15	0.07

Table 44: Provided are the growth, biomass and yield parameters of transgenic or control plants as measured in Green House under salinity irrigation. A = average; P = p value; RGR = Relative Growth Rate. The indicated days refer to days from planting.

TABLE 45

MAB79 - Salt irrigation (40-80 mM NaCl)

Event No.

7323.10,

7961.1,

7962.2,

	Control	A	P	A	P	A	P	A	P

Rosette Diameter on day 3	1.70			1.84	0.10	1.93	0.05
Rosette Diameter on day 5	2.36			2.53	0.01
Rosette Diameter on day 8	3.77			4.32	0.03	4.01	0.04
Rosette Area on day 3	0.90			1.08	0.09
Rosette Area on day 8	4.06			5.29	0.03	4.78	0.05
Plot Coverage on day 3	7.04			8.63	0.08
Plot Coverage on day 8	31.90			42.32	0.03	38.27	0.05
Leaf Number on day 3	5.08	5.63	0.00					5.75	0.04
Leaf Number on day 5	6.86							7.44	0.01
RGR of Leaf Number between	0.09			0.11	0.01
day 5 and 8
RGR of Rosette Area between	0.53							0.59	0.01
day 5 and 8
1000 Seeds weight [gr]	0.02			0.02	0.00
Harvest Index	0.11			0.19	0.00

Table 45: Provided are the growth, biomass and yield parameters of transgenic or control plants as measured in Green House under salinity irrigation.
A = average;
P = p value;
RGR = Relative Growth Rate.
The indicated days refer to days from planting.

Tables 46-58 depict analyses of plant parameters (as describe above) overexpressing the polynucleotides of the invention under the regulation of the 6669 promoter under Normal Growth conditions [Normal irrigation included NaCl Electrical conductivity (E.C.) of 1-2].

TABLE 46

MAB115 - Normal Growth Conditions

Event No.

8564.1

8565.1

8565.2

	Control	A	P	A	P	A	P

Rosette Diameter on day 3	1.67	1.80	0.09	1.75	0.04	1.95	0.04
Rosette Diameter on day 8	3.60			4.00	0.09
Rosette Area on day 5	1.54			1.70	0.09
Rosette Area on day 8	3.98			4.38	0.06
Plot Coverage on day 5	12.30			13.61	0.06
Plot Coverage on day 8	31.82			35.07	0.02
Leaf Number on day 3	5.25			5.94	0.00
Leaf Number on day 5	6.52			7.38	0.05
Leaf Number on day 8	8.92					9.31	0.00
RGR of Leaf Number between day 3 and 5	0.12			0.12	0.04
RGR of Rosette Area between day 5 and 8	0.53	0.62	0.01

Table 46: Provided are the growth, biomass and yield parameters of transgenic or control plants as measured in Green House under normal irrigation.
A = average;
P = p value;
RGR = Relative Growth Rate.
The indicated days refer to days from planting.

TABLE 47

MAB54 - Normal Growth Conditions

Event No.

8181.2

8182.2

8184.3

8185.4

	Control	A	P	A	P	A	P	A	P

Rosette Diameter on day 5	2.24	2.70	0.02	2.64	0.00			2.81	0.03
Rosette Diameter on day 8	3.60	4.00	0.08					4.29	0.05
Rosette Area on day 3	0.89			1.19	0.04			1.35	0.01
Rosette Area on day 8	3.98							5.91	0.05
Plot Coverage on day 3	7.10			9.52	0.04	7.49	0.08	10.76	0.01
Plot Coverage on day 8	31.82							47.28	0.06
Leaf Number on day 3	5.25	6.19	0.06					6.25	0.00
Leaf Number on day 5	6.52							7.94	0.03
Leaf Number on day 8	8.92	9.38	0.00					9.50	0.01
RGR of Leaf Number between day	0.12	0.13	0.08
3 and 5
RGR of Rosette Area between day	0.46	0.52	0.01					0.51	0.05
1 and 3
1000 Seeds weight [gr]	0.02			0.02	0.00	0.02	0.00

Table 47: Provided are the growth, biomass and yield parameters of transgenic or control plants as measured in Green House under normal irrigation.
A = average;
P = p value;
RGR = Relative Growth Rate.
The indicated days refer to days from planting.

TABLE 48

MAB55 - Normal Growth Conditions

Event No.

6802.5

6805.4

	Control	A	P	A	P

Rosette Area on day 5	1.54	1.70	0.03
Rosette Area on day 8	3.98			4.63	0.02
Plot Coverage on day 5	12.30	13.60	0.02
Plot Coverage on day 8	31.82			37.03	0.01

Table 48: Provided are the growth, biomass and yield parameters of transgenic or control plants as measured in Green House under normal irrigation. A = average; P = p value; RGR = Relative Growth Rate. The indicated days refer to days from planting.

TABLE 49

MAB56 - Normal Growth Conditions

Event No.

6691.2

6691.3

6693.2

6695.7

	Control	A	P	A	P	A	P	A	P

Rosette Diameter on day 3	1.67			1.89	0.05	1.97	0.07
Rosette Diameter on day 5	2.24			2.55	0.09	2.58	0.01
Rosette Area on day 3	0.89	1.06	0.00	1.15	0.02	1.23	0.08
Rosette Area on day 5	1.54			1.94	0.02	1.94	0.03
Plot Coverage on day 3	7.10	8.45	0.00	9.21	0.01	9.82	0.07
Plot Coverage on day 5	12.30			15.49	0.01	15.53	0.02
Plot Coverage on day 8	31.82					34.82	0.05
Leaf Number on day 3	5.25	5.50	0.00	5.81	0.00	6.00	0.02
Leaf Number on day 5	6.52			7.38	0.01	7.50	0.02
RGR of Leaf Number between	0.12					0.13	0.06
day 3 and 5
RGR of Leaf Number between	0.12							0.14	0.01
day 5 and 8
RGR of Rosette Area between	0.53							0.62	0.02
day 5 and 8
1000 Seeds weight [gr]	0.02	0.02	0.08

Table 49: Provided are the growth, biomass and yield parameters of transgenic or control plants as measured in Green House under normal irrigation.
A = average;
P = p value;
RGR = Relative Growth Rate.
The indicated days refer to days from planting.

TABLE 50

MAB57 - Normal Growth Conditions

Event No.

6912.1

6912.6

6912.9

	Control	A	P	A	P	A	P

Rosette Area on	3.98			4.48	0.02
day 8
Plot Coverage on	31.82			35.81	0.01
day 8
Leaf Number on	5.25					5.69	0.00
day 3
Leaf Number on	6.52	8.13	0.06	8.13	0.06
day 5
Leaf Number on	8.92					9.56	0.06
day 8
RGR of Rosette	0.53					0.58	0.10
Area between
day 5 and 8
1000 Seeds weight	0.02					0.02	0.00
[gr]

Table 50: Provided are the growth, biomass and yield parameters of transgenic or control plants as measured in Green House under normal irrigation.
A = average;
P = p value;
RGR = Relative Growth Rate.
The indicated days refer to days from planting.

TABLE 51

MAB58 - Normal Growth Conditions

Event No.

6783.2

7522.10

7522.3

7523.6

	Control	A	P	A	P	A	P	A	P

Rosette Diameter on day 3	1.67					2.11	0.00
Rosette Diameter on day 5	2.24	2.57	0.00
Rosette Diameter on day 8	3.60			4.06	0.07	4.54	0.00
Rosette Area on day 3	0.89					1.36	0.00
Rosette Area on day 5	1.54					2.29	0.00
Rosette Area on day 8	3.98					5.98	0.00
Plot Coverage on day 3	7.10					10.90	0.00
Plot Coverage on day 5	12.30					18.32	0.00
Plot Coverage on day 8	31.82					47.88	0.00
Leaf Number on day 3	5.25					6.06	0.00	6.44	0.05
Leaf Number on day 5	6.52							8.38	0.00
Leaf Number on day 8	8.92	9.25	0.04			9.81	0.04
RGR of Leaf Number between day	0.12	0.12	0.03
5 and 8
1000 Seeds weight [gr]	0.02	0.02	0.08

Table 51: Provided are the growth, biomass and yield parameters of transgenic or control plants as measured in Green House under normal irrigation.
A = average;
P = p value;
RGR = Relative Growth Rate.
The indicated days refer to days from planting.

TABLE 52

MAB59 - Normal Growth Conditions

Event No.

6793.4

6794.4

6794.5

	Control	A	P	A	P	A	P

Rosette Diameter on	1.67			2.38	0.05
day 3
Rosette Diameter on	2.24	2.73	0.06
day 5
Rosette Area on	0.89	1.27	0.06	1.65	0.03
day 3
Plot Coverage on	7.10	10.15	0.06	13.19	0.03
day 3
Leaf Number on	5.25	6.00	0.02	6.75	0.01
day 3
Leaf Number on	6.52	7.69	0.05	8.00	0.07
day 5
Leaf Number on	8.92			9.38	0.02
day 8
RGR of Rosette	0.46					0.49	0.02
Area between
day 1 and 3
1000 Seeds weight	0.02					0.02	0.00
[gr]

Table 52: Provided are the growth, biomass and yield parameters of transgenic or control plants as measured in Green House under normal irrigation.
A = average;
P = p value;
RGR = Relative Growth Rate.
The indicated days refer to days from planting.

TABLE 53

MAB69 - Normal Growth Conditions

Event No.

6651.11

6651.12

8341.1

8342.1

	Control	A	P	A	P	A	P	A	P

Rosette Diameter on day 3	1.67	1.92	0.01
Rosette Area on day 3	0.89	1.09	0.00
Rosette Area on day 8	3.98	5.05	0.04
Plot Coverage on day 3	7.10	8.74	0.00
Plot Coverage on day 8	31.82	40.41	0.04
Leaf Number on day 3	5.25	5.69	0.00					5.94	0.09
Leaf Number on day 8	8.92					8.94	0.08	9.31	0.00
RGR of Rosette Area between day	0.46	0.51	0.04
1 and 3
1000 Seeds weight [gr]	0.02			0.02	0.01

Table 53: Provided are the growth, biomass and yield parameters of transgenic or control plants as measured in Green House under normal irrigation.
A = average;
P = p value;
RGR = Relative Growth Rate.
The indicated days refer to days from planting.

TABLE 54

MAB70 - Normal Growth Conditions

Event No.

7971.3

7972.1

7974.3

	Control	A	P	A	P	A	P

Rosette Diameter on day 3	1.67			1.94	0.00	2.27	0.00
Rosette Diameter on day 5	2.24			2.62	0.04	3.03	0.00
Rosette Diameter on day 8	3.60	4.40	0.00			5.01	0.05
Rosette Area on day 3	0.89	1.20	0.07	1.12	0.01	1.52	0.04
Rosette Area on day 5	1.54	2.06	0.05	1.87	0.00	2.49	0.10
Rosette Area on day 8	3.98	5.86	0.06			7.03	0.05
Plot Coverage on day 3	7.10	9.61	0.06	9.00	0.01	12.12	0.04
Plot Coverage on day 5	12.30	16.46	0.04	14.93	0.00	19.90	0.09
Plot Coverage on day 8	31.82	46.87	0.06			56.23	0.05
Leaf Number on day 3	5.25					6.44	0.05
Leaf Number on day 5	6.52					8.13	0.00
Leaf Number on day 8	8.92			9.75	0.09	10.38	0.05
RGR of Rosette Area between day	0.53	0.62	0.01			0.61	0.03
5 and 8

Table 54: Provided are the growth, biomass and yield parameters of transgenic or control plants as measured in Green House under normal irrigation.
A = average;
P = p value;
RGR = Relative Growth Rate.
The indicated days refer to days from planting.

TABLE 55

MAB71 - Normal Growth Conditions

Event No.

7331.4

7332.2

7333.5

7334.5

	Control	A	P	A	P	A	P	A	P

Rosette Diameter on day 5	2.24			2.52	0.02
Rosette Diameter on day 8	3.60	4.33	0.01
Rosette Area on day 5	1.54							1.88	0.07
Rosette Area on day 8	3.98							4.68	0.00
Plot Coverage on day 5	12.30			14.25	0.08			15.05	0.05
Plot Coverage on day 8	31.82							37.48	0.00
Leaf Number on day 3	5.25			5.47	0.06			5.81	0.00
Leaf Number on day 5	6.52	7.56	0.00
RGR of Leaf Number between	0.16							0.17	0.01
day 1 and 3
RGR of Leaf Number between	0.12					0.12	0.10
day 3 and 5
RGR of Leaf Number between	0.12					0.12	0.01
day 5 and 8
RGR of Rosette Area between	0.53					0.61	0.04
day 5 and 8

Table 55: Provided are the growth, biomass and yield parameters of transgenic or control plants as measured in Green House under normal irrigation.
A = average;
P = p value;
RGR = Relative Growth Rate.
The indicated days refer to days from planting.

TABLE 54

MAB72 - Normal Growth Conditions

Event No.

8552.4

8553.2

8553.3

	Control	A	P	A	P	A	P

Rosette Diameter on	1.67	1.90	0.05			1.83	0.00
day 3
Rosette Diameter on	2.24	2.67	0.00			2.55	0.01
day 5
Rosette Diameter on	3.60	4.06	0.08	4.15	0.10
day 8
Rosette Area on	0.89	1.20	0.00	1.02	0.00	1.08	0.02
day 3
Rosette Area on	1.54	2.00	0.00			1.87	0.00
day 5
Rosette Area on	3.98	5.05	0.00			4.82	0.00
day 8
Plot Coverage on	7.10	9.64	0.00	8.13	0.00	8.67	0.02
day 3
Plot Coverage on	12.30	16.04	0.00			14.98	0.00
day 5
Plot Coverage on	31.82	40.39	0.00			38.57	0.00
day 8
Leaf Number on	5.25	5.88	0.00	5.56	0.00	5.50	0.00
day 3
Leaf Number on	8.92			9.38	0.02
day 8

Table 54: Provided are the growth, biomass and yield parameters of transgenic or control plants as measured in Green House under normal irrigation.
A = average;
P = p value;
RGR = Relative Growth Rate.
The indicated days refer to days from planting.

TABLE 55

MAB74 - Normal Growth Conditions

Event No.

7981.1

7982.4

7983.6

7983.9

	Control	A	P	A	P	A	P	A	P

Rosette Diameter on day 3	1.67			2.06	0.10	1.94	0.00
Rosette Diameter on day 5	2.24					2.62	0.06
Rosette Diameter on day 8	3.60					4.05	0.02
Rosette Area on day 3	0.89			1.14	0.02
Rosette Area on day 5	1.54			1.83	0.05	1.85	0.10
Rosette Area on day 8	3.98					4.46	0.03
Plot Coverage on day 3	7.10			9.13	0.02
Plot Coverage on day 5	12.30			14.62	0.03	14.79	0.08
Plot Coverage on day 8	31.82					35.66	0.01
Leaf Number on day 3	5.25			5.75	0.04	5.31	0.07
Leaf Number on day 5	6.52	6.26	0.02			7.25	0.04
Leaf Number on day 8	8.92							9.13	0.07
RGR of Leaf Number between	0.12			0.13	0.08			0.12	0.03
day 3 and 5
RGR of Rosette Area between	0.46							0.33	0.00
day 1 and 3
RGR of Rosette Area between	0.36	0.42	0.07
day 3 and 5
Biomass DW [gr]	3.07
1000 Seeds weight [gr]	0.02	0.02	0.02			0.02	0.05

Table 55: Provided are the growth, biomass and yield parameters of transgenic or control plants as measured in Green House under normal irrigation.
A = average;
P = p value;
RGR = Relative Growth Rate.
The indicated days refer to days from planting.

TABLE 56

MAB76 - Normal Growth Conditions

Event No.

7633.1

7635.16

	Control	A	P	A	P

Rosette Diameter on day 3	1.67			1.93	0.04
Leaf Number on day 3	5.25	5.44	0.02
Leaf Number on day 5	6.52			7.25	0.04

Table 56: Provided are the growth, biomass and yield parameters of transgenic or control plants as measured in Green House under normal irrigation. A = average; P = p value; RGR = Relative Growth Rate. The indicated days refer to days from planting.

TABLE 57

MAB77 - Normal Growth Conditions

Event No.

8212.1

8212.2

	Control	A	P	A	P

Leaf Number on day 3	5.25			5.63	0.00
Leaf Number on day 8	8.92	9.75	0.09

Table 57: Provided are the growth, biomass and yield parameters of transgenic or control plants as measured in Green House under normal irrigation. A = average; P = p value; RGR = Relative Growth Rate. The indicated days refer to days from planting.

TABLE 58

MAB79 - Normal Growth Conditions

Event No.

7324.1

7961.1

7962.2

	Control	A	P	A	P	A	P

Rosette Diameter on day 3	1.67	1.84	0.10	1.93	0.05
Rosette Diameter on day 5	2.24	2.53	0.01
Rosette Diameter on day 8	3.60	4.32	0.03	4.01	0.04
Rosette Area on day 3	0.89	1.08	0.09
Rosette Area on day 8	3.98	5.29	0.03	4.78	0.05
Plot Coverage on day 3	7.10	8.63	0.08
Plot Coverage on day 8	31.82	42.32	0.03	38.27	0.05
Leaf Number on day 3	5.25					5.75	0.04
Leaf Number on day 5	6.52					7.44	0.01
RGR of Rosette Area between day	0.53					0.59	0.01
5 and 8
1000 Seeds weight [gr]	0.02	0.02	0.00

Table 58: Provided are the growth, biomass and yield parameters of transgenic or control plants as measured in Green House under normal irrigation.
A = average;
P = p value;
RGR = Relative Growth Rate.
The indicated days refer to days from planting.

Example 7

Improved Fertilizer Efficiency in Arabidopsis Tissue Culture Assay

Plants transgenic to the following MAB genes were assayed for fertilizer use efficiency in a tissue culture assay: MAB115, MAB54, MAB55, MAB56, MAB57, MAB58, MAB59, MAB69, MAB70, MAB71, MAB72, MAB74, MAB76, MAB77, MAB79, MAB116, and MAB117 (the sequence identifiers of the cloned polynucleotides and their expressed polypeptides are provided in Table 7 above).
Assay I: Plant Growth at Nitrogen Deficiency Under Tissue Culture Conditions
The present inventors have found the nitrogen use efficiency (NUE) assay to be relevant for the evaluation of the ABST candidate genes, since NUE deficiency encourages root elongation, increase of root coverage and allows detecting the potential of the plant to generate a better root system under drought conditions. In addition, there are indications in the literature that biological mechanisms of NUE and drought tolerance are linked (Wesley et al., 2002 Journal of Experiment Botany Vol 53, No. 366, pp. 13-25).
Surface sterilized seeds were sown in basal media [50% Murashige-Skoog medium (MS) supplemented with 0.8% plant agar as solidifying agent] in the presence of Kanamycin (for selecting only transgenic plants). After sowing, plates were transferred for 2-3 days for stratification at 4° C. and then grown at 25° C. under 12-hour light 12-hour dark daily cycles for 7 to 10 days. At this time point, seedlings randomly chosen were carefully transferred to plates with nitrogen-limiting conditions: 0.5 MS media in which the combined nitrogen concentration (NH₄NO₃and KNO₃) is 0.75 mM (nitrogen deficient conditions) or 3 mM [Norman (optimal) nitrogen concentration]. Each plate contains 5 seedlings of same event, and 3-4 different plates (replicates) for each event. For each polynucleotide of the invention at least four independent transformation events were analyzed from each construct. Plants expressing the polynucleotides of the invention were compared to the average measurement of the control plants (empty vector or GUS reporter under the same promoter) used in the same experiment.
Digital imaging and statistical analysis—Parameters were measured and analyzed as previously described in Example 5, Assay 1 above.
Tables 59-68 depict analyses of seedling parameters (as describe above) overexpressing the polynucleotides of the invention under the regulation of At6669 promoter under Nitrogene Deficiency conditions.

TABLE 59

MAB70 - Nitrogene Deficiency (0.75 mM Nitrogen)

Event No.

7971.3

7972.1

7974.1

7974.3

	Control	A	P	A	P	A	P	A	P

Dry Weight [gr]	0.01							0.01	0.00
Fresh Wight [gr]	0.10							0.18	0.04
Leaf Area on day 10	0.36					0.47	0.04	0.55	0.00
Leaf Area on day 5	0.14							0.24	0.04
Roots Coverage on day 10	5.58	7.93	0.01	7.45	0.06
Roots Coverage on day 5	1.75	2.41	0.00					2.58	0.06
Roots Length on day 10	5.19	6.16	0.00
Roots Length on day 5	2.86	3.16	0.05
RGR of Roots Coverage between	0.45					0.67	0.05
day 5 and 10
RGR of Roots Coverage between	0.81	2.35	0.02	2.01	0.06	2.10	0.05	2.23	0.02
day 1 and 5
RGR of Roots Length between day	0.16					0.24	0.04
1 and 5
RGR of Roots Length between day	0.20	0.50	0.00	0.48	0.00	0.56	0.00	0.58	0.00
5 and 10

Table 59: Provided are the growth and biomass parameters of transgenic or control plants as measured in Tissue Calture growth under Nitrogene Deficiency (0.75 mM N).
A = average;
P = p value;
RGR = Relative Growth Rate.
The indicated days refer to days from planting.

TABLE 60

MAB71 - Nitrogene Deficiency (0.75 mM Nitrogen)

Event No.

7331.5

7332.2

7333.5

7334.4

	Control	A	P	A	P	A	P	A	P

Dry Weight [gr]	0.01	0.01	0.00	0.01	0.01	0.01	0.01	0.01	0.01
Fresh Wight [gr]	0.10	0.22	0.00	0.18	0.02	0.13	0.09
Leaf Area on day 10	0.36	0.55	0.00					0.52	0.00
Leaf Area on day 5	0.14	0.28	0.00					0.21	0.00
Roots Coverage on day 10	5.58					8.19	0.05	9.47	0.03
Roots Coverage on day 5	1.75	2.33	0.10					2.99	0.02
Roots Length on day 10	5.19							6.69	0.03
Roots Length on day 5	2.86							3.84	0.01
RGR of Roots Length between day	0.16			0.20	0.07
1 and 5
RGR of Roots Length between day	0.20	0.66	0.05			0.26	0.08
5 and 10

Table 60: Provided are the growth and biomass parameters of transgenic or control plants as measured in Tissue Calture growth under Nitrogene Deficiency (0.75 mM N).
A = average;
P = p value;
RGR = Relative Growth Rate.
The indicated days refer to days from planting.

TABLE 61

MAB74 - Nitrogene Deficiency (0.75 mM Nitrogen)

Event No.

7982.4

7983.9

	Control	A	P	A	P

Roots Coverage on day 10	5.58			9.76	0.06
Roots Length on day 10	5.19			6.70	0.00
RGR Leaf Area between	0.30			0.44	0.09
day 5 and 10
RGR of Roots Coverage	0.45	0.63	0.08
between day 5 and 10

Table 61: Provided are the growth and biomass parameters of transgenic or control plants as measured in Tissue Calture growth under Nitrogene Deficiency (0.75 mM N). A = average; P = p value; RGR = Relative Growth Rate. The indicated days refer to days from planting.

TABLE 62

MAB76 - Nitrogene Deficiency (0.75 mM Nitrogen)

Event No.

7635.4

	Control	A	P

RGR of Roots Coverage	0.45	0.70	0.07
between day 5 and 10
RGR of Roots Length	0.16	0.26	0.07
between day 1 and 5

Table 62: Provided are the growth and biomass parameters of transgenic or control plants as measured in Tissue Calture growth under Nitrogene Deficiency (0.75 mM N). A = average; P = p value; RGR = Relative Growth Rate. The indicated days refer to days from planting.

TABLE 63

MAB77 - Nitrogene Deficiency (0.75 mM Nitrogen)

Event No.

7931.11

8211.8

8212.2

	Control	A	P	A	P	A	P

Dry Weight [gr]	0.01					0.01	0.00
Fresh Wight [gr]	0.10			0.13	0.03	0.14	0.01
Roots Coverage on	1.75	2.26	0.03
day 5
RGR of Roots	0.45			0.71	0.01	0.75	0.05
Coverage between
day 5 and 10
RGR of Roots	0.16			0.24	0.03
Length between
day 1 and 5
RGR of Roots	0.20	0.31	0.05	0.99	0.04	0.86	0.08
Length between
day 5 and 10

Table 63: Provided are the growth and biomass parameters of transgenic or control plants as measured in Tissue Calture growth under Nitrogene Deficiency (0.75 mM N).
A = average;
P = p value;
RGR = Relative Growth Rate.
The indicated days refer to days from planting.

TABLE 64

MAB79 - Nitrogene Deficiency (0.75 mM Nitrogen)

Event No.

7323.3

7324.1

7961.1

	Control	A	P	A	P	A	P

Dry Weight [gr]	0.01	0.01	0.00	0.01	0.03	0.01	0.01
Fresh Wight [gr]	0.10	0.16	0.00			0.11	0.10
RGR of Roots	0.45	0.71	0.09
Coverage between
day 5 and 10
RGR of Roots	0.81	3.11	0.00
Coverage between
day 1 and 5
RGR of Roots	0.20	0.67	0.00	0.49	0.09
Length between
day 5 and 10

Table 64: Provided are the growth and biomass parameters of transgenic or control plants as measured in Tissue Calture growth under Nitrogene Deficiency (0.75 mM N).
A = average;
P = p value;
RGR = Relative Growth Rate.
The indicated days refer to days from planting.

TABLE 65

MAB115 - Nitrogene Deficiency (0.75 mM Nitrogen)

Event No.

8561.2

8564.2

8565.1

	Control	A	P	A	P	A	P

Dry Weight [gr]	0.005	0.006	0.00	0.010	0.00	0.009	0.00
Leaf Area on day 10	0.24					0.27	0.00
Leaf Area on day 5	0.35					0.38	0.01
Roots Coverage on day 10	1.67					2.13	0.02
Roots Coverage on day 5	3.38					4.80	0.03
Roots Length on day 10	2.39					2.74	0.00
Roots Length on day 5	3.46					4.22	0.02
RGR Leaf Area between day 5 and 10	0.37	0.43	0.00			0.48	0.00
RGR Leaf Area between day 1 and 5	0.16	0.19	0.00
RGR of Roots Coverage between day 5 and 10	1.89	3.21	0.00	2.23	0.02	3.47	0.01
RGR of Roots Coverage between day 1 and 5	0.33	0.35	0.00	0.43	0.00	0.44	0.00
RGR of Roots Length between day 1 and 5	0.37	0.56	0.02	0.53	0.06	0.68	0.00
RGR of Roots Length between day 5 and 10	0.15			0.17	0.00	0.18	0.00

Table 65: Provided are the growth and biomass parameters of transgenic or control plants as measured in Tissue Calture growth under Nitrogene Deficiency (0.75 mM N).
A = average;
P = p value;
RGR = Relative Growth Rate.
The indicated days refer to days from planting.

TABLE 66

MAB54 - Nitrogene Deficiency (0.75 mM Nitrogen)

Event No.

8181.2

8182.2

8185.3

	Control	A	P	A	P	A	P

Dry Weight [gr]	0.005	0.009	0.00	0.008	0.00	0.007	0.00
Roots Coverage on day 10	1.67	1.80	0.04
Roots Coverage on day 5	3.38	4.27	0.02	3.40	0.00
Roots Length on day 10	2.39	2.52	0.01
Roots Length on day 5	3.46	4.11	0.02	3.50	0.00
RGR Leaf Area between day 5 and 10	0.37	0.45	0.00
RGR Leaf Area between day 1 and 5	0.16	0.17	0.04			0.19	0.00
RGR of Roots Coverage between day 5 and 10	1.89	3.59	0.00	3.60	0.01	2.20	0.01
RGR of Roots Coverage between day 1 and 5	0.33	0.52	0.00	0.55	0.00	0.36	0.00
RGR of Roots Length between day 1 and 5	0.37	0.70	0.00	0.73	0.00	0.51	0.02
RGR of Roots Length between day 5 and 10	0.15	0.21	0.01	0.22	0.01	0.17	0.00

Table 66: Provided are the growth and biomass parameters of transgenic or control plants as measured in Tissue Calture growth under Nitrogene Deficiency (0.75 mM N).
A = average;
P = p value;
RGR = Relative Growth Rate.
The indicated days refer to days from planting.

TABLE 67

MAB57 - Nitrogene Deficiency (0.75 mM Nitrogen)

Event No.

6912.14

6912.20

6912.60

	Control	A	P	A	P	A	P

Roots Coverage on	5.55			6.71	0.10
day 7
Roots Coverage	15.02	17.33	0.05
on day 14
Roots Length on	4.93	5.54	0.10	6.12	0.02	6.34	0.02
day 7
Roots Length on	7.83	8.76	0.02
day 14
Fresh Weight	0.19	0.24	0.09

Table 67: Provided are the growth and biomass parameters of transgenic or control plants as measured in Tissue Calture growth under Nitrogene Deficiency (0.75 mM N).
A = average;
P = p value;
RGR = Relative Growth Rate.
The indicated days refer to days from planting.

TABLE 68

MAB72 - Nitrogene Deficiency (0.75 mM Nitrogen)

Event No.

8552.1

8552.4

8553.2

	Control	A	P	A	P	A	P

Dry Weight [gr]	0.005	0.01	0.00	0.01	0.00	0.01	0.00
Leaf Area on day 10	0.24	0.24	0.01
Roots Coverage on day 10	1.67	1.84	0.01	1.93	0.02	1.89	0.03
Roots Coverage on day 5	3.38	3.60	0.04	3.90	0.06	4.50	0.00
Roots Length on day 10	2.39	2.48	0.01
Roots Length on day 5	3.46	3.70	0.04	3.65	0.04	3.84	0.00
RGR Leaf Area between day 5 and 10	0.37	0.47	0.00	0.39	0.00
RGR Leaf Area between day 1 and 5	0.16			0.20	0.09
RGR of Roots Coverage between day 5 and 10	1.89	1.93	0.03	2.29	0.06	3.04	0.01
RGR of Roots Coverage between day 1 and 5	0.33			0.35	0.00	0.52	0.00
RGR of Roots Length between day 1 and 5	0.37					0.55	0.01
RGR of Roots Length between day 5 and 10	0.15	0.16	0.00	0.18	0.00	0.22	0.01

Table 68: Provided are the growth and biomass parameters of transgenic or control plants as measured in Tissue Calture growth under Nitrogene Deficiency (0.75 mM N).
A = average;
P = p value;
RGR = Relative Growth Rate.
The indicated days refer to days from planting.

Example 8

Transgenic Tomato and Arabidopsis Plants Show Improved Tolerance to Salt and Water-Deficiency Stresses Under Field Conditions

To test the impact of AQP TIP2 genes on plant's stress tolerance, the present inventors have previously cloned and overexpressed a polynucleotide which comprises the nucleic acid sequence set forth by SEQ ID NO:2827 (also known as ABST36 set forth by SEQ ID NO:13 in WO2004/104162; or S1TIP2;2) and which encodes the TIP2 polypeptide set forth by SEQ ID NO:2828 (which comprises the consensus sequence TLXFXFAGVGS; SEQ ID NO:2826). The nucleic acid constructs which comprises the ABST36 polynucleotide under the regulation of the constitutive Arabidopsis At6669 promoter (SEQ ID NO: 2823) (further referred to as the At6669::ABST36 construct) was further transformed into tomato (Solanum lycopersicum) as a model crop plant (Tom-ABST36), as well as into Arabidopsis thaliana. Four independent, T₂transgenic tomato genotypes, overexpressing ABST36 in heterozygous form, were evaluated for their tolerance to salt and water deficiency in two different salt-stress field trials and one water-deficiency-stress field trial consisting of two water-deficiency regimes. Transgenic genotypes in each field trial were compared to their null-segregant counterparts as controls.
Materials and Experimental Methods
Tomato Salt-stress field trial—All field trials were performed in a light soil, in an open field (net-house) near Rehovot, Israel. The F1 hybrids of four independent events of the cross between ABST36-transgenic MicroTom plants and M82 tomato plants were grown for the first 3 weeks in a nursery under normal irrigation conditions. The seedlings were then transplanted into rows and grown in a commercial greenhouse. The salt-stress trial was divided into four blocks. In each block, two different irrigation systems were established: a normal water regime for tomato cultivation and a continuous irrigation with saline water (addition of 180 to 200 mM NaCl). Each block consisted of a total of 60 plants divided as follows: six plants per event and six seedling null segregants were planted in the control row and a similar number of plants were planted in the salt-stressed row. At the stage of about 80% red fruits in planta, fruit yield, plant fresh weight, and harvest index were calculated. Harvest index was calculated as yield/plant biomass.
Tomato Water-deficiency-stress field trial—All field trials were performed in a light soil, in an open field (net-house) near Rehovot, Israel. The F1 hybrids of the four independent events were initially grown as described above. Three-week-old seedlings were transplanted to a net-greenhouse. The experiment was structured in four blocks containing three rows irrigated with different water levels and intervals (WLI-0, WLI-1, WLI-2). In each block, six transgenic plants per event analyzed and six non transgenic plants were transplanted in each row. Seedlings were transplanted after 4 weeks into wet soil. The amount of water used to uniformly irrigate before transplanting reached maximum water capacity [20% weight per weight (w/w)] at 60 cm depth, but without the creation of water overload. Each plant was transplanted near a dripper, with a 30-cm distance between plants, giving a total density of 2,600 plants per 1,000 m², according to a commercial growth protocol. Soil water capacity was measured using the standard procedures by sampling soil from the following three depths: 0 to 20 cm, 20 to 40 cm, and 40 to 60 cm. The water content in these soil layers was measured routinely every week. The soil contained 5% hygroscopic water while the maximum water capacity of the soil was 20%. All fertilizers were applied in the soil prior to plant transplantation. The amount of both phosphorus and potassium was calculated to be sufficient for all seasons. Nitrogen was applied as recommended, equally to all treatments, through the irrigation system. Each row contained three dripping irrigation lines creating a coverage of nine drippers per 1 m². The water control was performed separately for each treatment. The soil was dried completely before the beginning of the experiment. The different water regimes were begun only 4 weeks after transplanting when plants initiated the flowering stage. The amount of water supplied every week during the assay was calculated at the beginning of every week following the recommendations of standard growth protocols. WLI-0 treatment (control) received the recommended total weekly irrigation volume divided into three irrigations. WLI-1 was irrigated three times a week, but the amount of water supplied was half that supplied to WLI-0. At the end of every week, WLI-1 plants received the amount of water required to reach maximum soil water capacity. WLI-2 plants were irrigated only once a week, at the beginning of the week. The water-stress experiment lasted throughout the flowering period (23 days), corresponding to four cycles of the above-described stresses. Afterwards, all treatments received the recommended amount of water. The calculated water amount was equal to the difference between the water contents in dry soil and in soil with maximum water capacity. At the end of each stress cycle, the water amounts were compared between treatments according to actual water content in the soil (S3). During the stress period, treatments WLI-1 and WLI-2 received a total of 75% less water than the controls (WLI-0).
Experimental Results
Transgenic plants exhibit increased tolerance to salt stress—To induce salt-stress, transgenic and control tomato plants were continuously irrigated in field trials with 180 to 200 mM NaCl. As shown in FIGS. 3 a-c, 3 g-j and Table 69 below, Tom-ABST36 plants appeared to be more vigorous in all of the experiments than the control plants, which were smaller and showed severe symptoms of leaf and shoot necrosis (see for example, FIG. 3 j). This was also associated with higher fruit yield in Tom-ABST36 plants relative to controls (FIG. 3 a).

TABLE 69

Salt-stress field trial

Control

180 mM NaCl

Plant FW	Fruit yield	Harvest	Plant FW	Fruit yield		Harvest
(tn/acre)	(tn/acre)	index	(tn/acre)	(tn/acre)	%*	index

SlTIP2;2	ND	24.0	ND	2.8^a	8.0^a	110%	2.8
WT	ND	24.0	ND	1.4^b	3.8^b	0%	2.7

Table 69: Total yield (ton fruit/acre), plant fresh weight (FW), and harvest index were calculated for TOM ABST36 vs. control plants growing in the field under salt-stress conditions (180 mM NaCl). Results are the average of four independent events.
^a,bValues in a column followed by different superscript letters are significantly different.

Transgenic plants exhibit increased tolerance to water-deficiency stress—Transgenic plants subjected to water-deficiency stress exhibited a significantly higher (26%, p<0.05) plant biomass compared to control plants (FIG. 3 e). Moreover the Tom-ABST36 plants showed a significant (up to 21%, p≦0.05) increment of fruit yield under water-deficient regimes (water level intervals WLI-1), while under normal irrigation, the yield improvement was even higher (27%, p≦0.05; FIG. 3 d). The harvest index of the Tom-ABST36 plants was also higher when plants grew under regular and WLI-1 conditions while it remained similar to control when the water-deficient regime consisted of once-a-week irrigation (WLI-2) (FIG. 3 f).
The results from the three field trials provided strong evidence that the tomato Tom-ABST36 plants show improved tolerance to salt and water-deficiency stress relative to the control plants, which is translated into significant increments in plant biomass and more importantly, fruit yield.
Arabidopsis Salt-stress green house trial—A complementary experiment with transgenic Arabidopsis plants expressing the ABST36 construct showed increased tolerance to a salt stress of 150 mM NaCl compared to control plants, as reflected in 42% higher fresh biomass and 60% higher dry biomass (Table 70 below).
In-vitro salt-stress assay—Seeds of transgenic Arabidopsis plants harboring the At6669::ABST36 construct or 35S::GUS construct (which was used as control) were sown in 1/2 MS media containing 40 mg/l kanamycin for selection. Selected seedlings were sub-cultured to 1/2 MS media with 0 or 150 mM NaCl. Plants were grown for a period of 3 weeks. Results are the average of four independent events that were analyzed in four repeats. For the determination of shoot dry weight, shoot plants were collected and dried for 24 hours at 60° C. and then weighed.

TABLE 70

Arabidopsis salt-stress assay

0 mM NaCl

150 mM NaCl

	Plant FW	Plant DW	Plant FW	Plant DW
Lines	(mg)	(mg)	(mg)	(mg)

SlTIP2;2	408.28^a	23.52^a	68.55^a	4.4^a
WT	394.36^a	22.63^a	48.12^b	2.7^b

Table 70. Arabidopsis seedlings were grown in 0 and 150 mM NaCl under tissue-culture conditions. Shown are the fresh weight (FW) and total dry weight (DW) (both measured in milligrams) of ABST36 (SEQ ID NO: 2827) transgenic or wild type controls under normal conditions (0 mM NaCl) or salinity stress (150 mM NaCl).
^a,bValues in a column followed by different superscript letters are significantly different at P < 0.05

Although the invention has been described in conjunction with specific embodiments thereof, it is evident that many alternatives, modifications and variations will be apparent to those skilled in the art. Accordingly, it is intended to embrace all such alternatives, modifications and variations that fall within the spirit and broad scope of the appended claims.
All publications, patents and patent applications mentioned in this specification are herein incorporated in their entirety by reference into the specification, to the same extent as if each individual publication, patent or patent application was specifically and individually indicated to be incorporated herein by reference. In addition, citation or identification of any reference in this application shall not be construed as an admission that such reference is available as prior art to the present invention. To the extent that section headings are used, they should not be construed as necessarily limiting.

CD-ROM Content

The following lists the file content of the CD-ROM which is enclosed herewith and filed with the application. File information is provided as: File name/byte size/date of creation/operating system/machine format.
CD-ROM1 (1 file of SEQUENCE LISTING):

1. 44910_ST25.txt″/6,002,873 bytes/23 Dec. 2008/Microsoft Windows XP Professional/PC

Claims

1. A method of increasing abiotic stress tolerance, water use efficiency (WUE), fertilizer use efficiency (FUE), biomass, vigor and/or yield of a plant, comprising expressing within the plant an exogenous polynucleotide encoding a polypeptide comprising an amino acid sequence at least 80% homologous to the amino acid sequence selected from the group consisting of SEQ ID NOs: 2767, 33, 34, 30, 27-29, 31, 32, 35-52, 1401-1403, 1405-1435, 1437-1494, 1496-1542, 1544-1553, 1555-1559, 1561-1827, 1829-1866, 1868-2450, 2453-2458, 2460-2463, 2465-2481, 2483, 2485-2746, 2765-2766, 2768-2769, 3052-3065 and 3067-3259, thereby increasing the abiotic stress tolerance, water use efficiency (WUE), fertilizer use efficiency (FUE), biomass, vigor and/or yield of the plant.

2. A method of increasing abiotic stress tolerance, water use efficiency (WUE), fertilizer use efficiency (FUE), biomass, vigor and/or yield of a plant, comprising expressing within the plant an exogenous polynucleotide comprising a nucleic acid sequence at least 80% identical to the nucleic acid sequence selected from the group consisting of SEQ ID NOs: 2756, 7, 8, 4, 1-3, 5, 6, 9-26, 53-55, 57-87, 89-147, 149-195, 197-206, 208-212, 214-480, 482-519, 521-1103, 1106-1111, 1113-1116, 1118-1134, 1136, 1138-1400, 2748-2755, 2757-2764, 2843-2857 and 2859-3051, thereby increasing the abiotic stress tolerance, water use efficiency (WUE), the fertilizer use efficiency (FUE), the biomass, the vigor and/or the yield of the plant.

3. An isolated polynucleotide comprising a nucleic acid sequence at least 80% identical to the nucleic acid sequence selected from the group consisting of SEQ ID NOs: 2756, 7, 8, 4, 1-3, 5, 6, 9-26, 53-55, 57-87, 89-147, 149-195, 197-206, 208-212, 214-480, 482-519, 521-1103, 1106-1111, 1113-1116, 1118-1134, 1136, 1138-1400, 2748-2755, 2757-2764, 2843-2857 and 2859-3051.

4. A nucleic acid construct, comprising the isolated polynucleotide of claim 3 and a promoter for directing transcription of said nucleic acid sequence.

5. An isolated polypeptide, comprising an amino acid sequence at least 80% homologous to the amino acid sequence selected from the group consisting of SEQ ID NOs:2767 33, 34, 30, 27-29, 31, 32, 35-52, 1401-1403, 1405-1435, 1437-1494, 1496-1542, 1544-1553, 1555-1559, 1561-1827, 1829-1866, 1868-2450, 2453-2458, 2460-2463, 2465-2481, 2483, 2485-2746, 2765-2766, 2768-2769, 3052-3065 and 3067-3259.

6. A plant cell exogenously expressing the isolated polypeptide of claim 5.

7. A plant cell exogenously expressing the isolated polynucleotide of claim 3.

8. The method of claim 2, wherein said polynucleotide is selected from the group consisting of SEQ ID NOs:2756 7, 8, 4, 1-3, 5, 6, 9-26, 53-55, 57-87, 89-147, 149-195, 197-206, 208-212, 214-480, 482-519, 521-1103, 1106-1111, 1113-1116, 1118-1134, 1136, 1138-1400, 2748-2755, 2757-2764, 2843-2857 and 2859-3051.

9. The method of claim 1, wherein said amino acid sequence is selected from the group consisting of SEQ ID NOs:2767 33, 34, 30, 27-29, 31, 32, 35-52, 1401-1403, 1405-1435, 1437-1494, 1496-1542, 1544-1553, 1555-1559, 1561-1827, 1829-1866, 1868-2450, 2453-2458, 2460-2463, 2465-2481, 2483, 2485-2746, 2765-2766, 2768-2769, 3052-3065 and 3067-3259.

10. The method of claim 1, wherein said polypeptide is selected from the group consisting of SEQ ID NOs: 2767, 33, 34, 30, 27-29, 31, 32, 35-52, 1401-1403, 1405-1435, 1437-1494, 1496-1542, 1544-1553, 1555-1559, 1561-1827, 1829-1866, 1868-2450, 2453-2458, 2460-2463, 2465-2481, 2483, 2485-2746, 2765-2766, 2768-2769, 3052-3065 and 3067-3259.

11-12. (canceled)

13. The nucleic acid construct of claim 4, wherein said isolated polynucleotide comprising the nucleic acid sequence set forth by SEQ ID NO:2756 7, 8, 4, 1-3, 5, 6, 9-26, 53-55, 57-87, 89-147, 149-195, 197-206, 208-212, 214-480, 482-519, 521-1103, 1106-1111, 1113-1116, 1118-1134, 1136, 1138-1400, 2748-2755, 2757-2764, 2843-2857, 2859-3050 or 3051.

14. (canceled)

15. The isolated polypeptide of claim 5, wherein said amino acid sequence is set forth by SEQ ID NO: 2767, 33, 34, 30, 27-29, 31, 32, 35-52, 1401-1403, 1405-1435, 1437-1494, 1496-1542, 1544-1553, 1555-1559, 1561-1827, 1829-1866, 1868-2450, 2453-2458, 2460-2463, 2465-2481, 2483, 2485-2746, 2765-2766, 2768-2769, 3052-3065, 3067-3258 or 3259.

16. The plant cell of claim 6, wherein said polypeptide having the amino acid sequence set forth by SEQ ID NO: 2767, 33, 34, 30, 27-29, 31, 32, 35-52, 1401-1403, 1405-1435, 1437-1494, 1496-1542, 1544-1553, 1555-1559, 1561-1827, 1829-1866, 1868-2450, 2453-2458, 2460-2463, 2465-2481, 2483, 2485-2746, 2765-2766, 2768-2769, 3052-3065, 3067-3258 or 3259.

17. The plant cell of claim 7, wherein said polynucleotide comprising the nucleic acid sequence set forth by SEQ ID NO: 2756, 7, 8, 4, 1-3, 5, 6, 9-26, 53-55, 57-87, 89-147, 149-195, 197-206, 208-212, 214-480, 482-519, 521-1103, 1106-1111, 1113-1116, 1118-1134, 1136, 1138-1400, 2748-2755, 2757-2764, 2843-2857, 2859-3050 or 3051.

18. The method of claim 1, wherein the abiotic stress is selected from the group consisting of salinity, water deprivation, low temperature, high temperature, heavy metal toxicity, anaerobiosis, nutrient deficiency, nutrient excess, atmospheric pollution and UV irradiation.

19. The method of claim 1, further comprising growing the plant expressing said exogenous polynucleotide under the abiotic stress.

20. The nucleic acid construct of claim 4, wherein said promoter is a constitutive promoter.

21. The plant cell of claim 6, wherein said plant cell forms a part of a plant.

22. The plant cell of claim 7, wherein said plant cell forms a part of a plant.

23. The isolated polynucleotide of claim 3, wherein said polynucleotide is selected from the group consisting of SEQ ID NOs:2756, 7, 8, 4, 1-3, 5, 6, 9-26, 53-55, 57-87, 89-147, 149-195, 197-206, 208-212, 214-480, 482-519, 521-1103, 1106-1111, 1113-1116, 1118-1134, 1136, 1138-1400, 2748-2755, 2757-2764, 2843-2857 and 2859-3051.

24. The method of claim 1, wherein the plant is a dicotyledonous plant.

25. The method of claim 1, wherein the plant is a monocotyledonous plant.

26. The method of claim 2, wherein the plant is a dicotyledonous plant.

27. The method of claim 2, wherein the plant is a monocotyledonous plant.

28. The plant cell of claim 21, wherein the plant is a dicotyledonous plant.

29. The plant cell of claim 21, wherein the plant is a monocotyledonous plant.

30. A transgenic plant exogenously expressing the isolated polynucleotide of claim 3.

31. A transgenic plant transformed with the nucleic acid construct of claim 4.